Compositions containing non-polar compounds

ABSTRACT

Provided are compositions and methods for clear and stable beverages that contain additives such as essential fatty acids, including omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids, and other fatty acids; phytochemicals, including phytosterols; other oils; and coenzymes, including Coenzyme Q10, and other oil-based additives.

RELATED APPLICATIONS

This application is related to International PCT Application No.PCT/US2011/000538, filed Mar. 23, 2011, entitled “COMPOSITIONSCONTAINING NON-POLAR COMPOUNDS,” which also claims priority to U.S.Provisional Application Ser. No. 61/340,944, filed Mar. 23, 2010.

This application is related to U.S. application Ser. No. 12/383,244,filed Mar. 20, 2009, published as US-2009-0297665-A1, and entitled“COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS,” and InternationalApplication No. PCT/US2009/001775, filed Mar. 20, 2009, published asInternational PCT Application No. WO 2009/117152 and entitled “EMULSIONSINCLUDING A PEG-DERIVATIVE OF TOCOPHEROL,” all of which claim priorityto U.S. Provisional Application Ser. No. 61/070,381, filed Mar. 20,2008, entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS” and U.S.Provisional Application Ser. No. 61/132,424, filed Jun. 16, 2008,entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS,” each to PhilipBromley.

This application also is related to U.S. patent application Ser. No.12/383,241, filed Mar. 20, 2009, published as US-2009-0297491-A1entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS” and InternationalApplication No. PCT/US2009/001774, filed Mar. 20, 2009, published asInternational PCT Application No. WO 2009/117151 and entitled “VITAMIN EDERIVATIVES AND THEIR USES,” all of which claim priority to U.S.Provisional Application Ser. No. 61/070,392, filed Mar. 20, 2008,entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS” and U.S.Provisional Application Ser. No. 61/132,409, filed Jun. 16, 2008,entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS” each to PhilipBromley.

The subject matter of each of the above-referenced applications isincorporated by reference in its entirety.

FIELD OF THE INVENTION

Provided are compositions and methods for preparing foods and beveragesthat contain additives, such as nutraceuticals, pharmaceuticals andsupplements.

BACKGROUND

Non-polar compounds are not easily dissolved in aqueous solutions, suchas water or other polar solvents. A number of non-polar compounds areused in compositions for human ingestion, for example, pharmaceuticals,nutraceuticals and/or dietary supplements. Exemplary non-polar compoundsused in such compositions are vitamins and minerals, fatty acids, andother non-polar compounds, non-polar active agents, and non-polar activeingredients.

Because of poor water solubility, inclusion of non-polar compounds inproducts for human consumption, for example, supplements, foods andbeverages, often is challenging. Available compositions containingnon-polar compounds, particularly aqueous compositions containingnon-polar compounds, and methods for formulating such compositions, arelimited. Thus, there remains a need to develop compositions containingnon-polar compounds and methods for making the compositions.Accordingly, it is among the objects herein to provide compositions,including aqueous compositions, containing non-polar compounds, andmethods for making the compositions.

SUMMARY

Provided are first compositions (concentrates) that contain non-polarcompounds, including liquid nanoemulsion concentrates. Also provided aremethods that use such first compositions to prepare other compositions,such as beverages and other aqueous liquids, into which the firstcompositions are diluted to form liquid dilution compositions. Alsoprovided are liquid dilution compositions containing the beverage orother aqueous liquid and the diluted concentrate. The concentratescontain dispersions, and/or can be used to prepare dispersions, ofeffective amounts of additives, such as non-polar compounds, includingnon-polar active ingredients, such as nutraceuticals, pharmaceuticals,and supplements, such as essential fatty acids, includingpolyunsaturated fatty acids, such as omega-3 fatty acids, omega-6 fattyacids, conjugated fatty acids, and other fatty acids; phytochemicals,including phytosterols; other oils; coenzymes, including Coenzyme Q10;vitamins, including vitamin D3; and other oil-based additives. Theamounts in the resulting diluted compositions are effective tosupplement the diet. The compositions provided herein are stabledispersions without phase separation or other changes.

For example, the provided compositions include concentrates containingnon-polar active ingredients, surfactants, and polar solvents, atamounts whereby dilution of the concentrate in an aqueous medium, suchas a beverage, at a particular amount (e.g., any of the specifiedamounts, concentrations, and dilutions of the concentrates and any ofthe amounts of the non-polar active ingredients described herein below),yields a liquid dilution composition containing effective amounts of thenon-polar active ingredient and having one or more desired properties.The provided compositions further include liquid dilution compositions,including the liquid dilution compositions made from the concentrates,containing aqueous media, non-polar active ingredients at effectiveamounts and polar solvents that have the desired properties. The amountof the concentrate and/or amount of the non-polar active ingredient canbe specified. The desired properties include clarity of the liquiddilution compositions, such as compositions that are clear or about asclear as the aqueous medium in the absence of the concentrate and/or inthe absence of the non-polar active ingredient; particle size, such asparticle size of less than 200 nm or less than about 200 nm, less than100 nm or less than about 100 nm, less than 50 nm or less than about 50nm, or less than 25 nm or less than about 25 nm, at most or on average;turbidity, such as a Nephelometric Turbidity Units (NTU) value of lessthan 200 or about 200, less than 100 or about 100, less than 50 or about50, less than 30 or about 30, less than 25 or about 25, or less than 10or about 10; and the lack of visible particles, visible crystals, phaseseparation, and/or ringing.

The provided concentrates typically are liquid nanoemulsionconcentrates, which contain surfactants (typically a surfactant that isa sugar fatty acid ester or mixture of sugar fatty acids esters, suchas, for example a sucrose fatty acid ester or a mixture of sucrose fattyacids esters or a mixture of the sucrose fatty acid esters and aPEG-derivative of Vitamin E), non-polar compound(s) (which typicallyis/are a non-polar active ingredient which differs from the surfactant)and a polar solvent (e.g., water or other edible aqueous liquid, such asa polar protic solvent such as a dihydric or trihydric alcohol, e.g.,propylene glycol and glycerin (glycerol)).

The amount of non-polar compound in the concentrate typically is between5% or about 5% and 10% or about 10%, by weight (w/w), of theconcentrate, e.g., at or about 5, 5.2, 5.25, 6, 7, 8, 9, or 10%, byweight, of the concentrate.

The surfactants in the provided concentrates typically have aHydrophilic Lipophilic Balance (HLB) value of between 14 or about 14 and20 or about 20, such as between 15 or about 15 and 18 or about 18, e.g.,at or about 15, 16, 17, or 18. Exemplary surfactants for use in thecompositions provided include non-ionic surfactants, such as sugar estersurfactants, such as sugar fatty acid ester surfactants, typically,sucrose fatty acid ester surfactants, which typically contain sucrosefatty acid monoesters (e.g., sucrose fatty acid ester surfactants).

The amount of surfactant(s) in the concentrate typically is between 16%or about 16% and 30% or about 30%, by weight, of the concentrate, e.g.,at or about 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or30%, by weight (w/w), of the concentrate. When the concentrate containsa mixture of sucrose fatty acid esters and a PEG-derivative of Vitamin Eas surfactants, the compositions typically contain at least 1% or atleast about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% sucrose fattyesters.

In some examples, the amount of the surfactant(s) in the concentrate isbetween 17% or about 17% and 26% or about 26%, by weight (w/w), of theconcentrate, for example, between 18% or about 18% and 26% or about 26%,or between 16% or about 16% and 18% or about 18%. In some examples, theamount of the surfactant(s) in the concentrate is at or about 16, 17,18, 19, 20, 21, 22, 23, 24, 25, or 26%, by weight (w/w), of theconcentrate, such as, for example, 17.75%, 20.25%, 20.5%, 22.7%, or25.2% (w/w) of the concentrate.

The amount of polar solvent in the concentrate typically is between 60%or about 60% and 79% or about 79%, by weight (w/w), of the concentrate,for example, at or about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, or 79%, by weight (w/w), of the concentrate.

In one example, the amount of polar solvent in the concentrate isbetween 65% or about 65% and 79% or about 79%, between 65% or about 65%and 75% or about 75%, between 65% or about 65% and 76% or about 76%,between 68% or about 68% and 76% or about 76%, by weight (w/w), of theconcentrate. For example, the amount of polar solvent in the concentratecan be at or about 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, or 80%, by weight (w/w), of the concentrate, such as at or about74.25%, at or about 75.8%, at or about 68.9%, at or about 71.74%, at orabout 63.94%, at or about 68.79%, at or about 68.29%, at or about69.02%, or at or about 71.49%, by weight (w/w), of the concentrate.

Among the sugar ester surfactants are sugar fatty acid estersurfactants, typically sucrose fatty acid ester surfactants. The sugarfatty acid ester surfactants can be blends of different esters, such assugar fatty acid monoesters, diesters, triesters, and polyesters. Thesucrose fatty acid ester surfactants typically contain sucrose fattyacid monoesters. The sucrose fatty acid esters containing monoestersfurther can contain one or more of sucrose fatty acid diesters,triesters, and/or polyesters. In one example, the sucrose fatty acidester contains one or more of sucrose stearate, sucrose laurate, sucrosepalmitate, sucrose oleate, sucrose caprylate, sucrose decanoate, sucrosemyristate, sucrose pelargonate, sucrose undecanoate, sucrosetridecanoate, sucrose pentadeconoate, sucrose heptadecanoate, andhomologs thereof, including mono-, di-, tri-, and poly-ester forms ofthese sucrose fatty acid esters.

The sucrose fatty acid ester surfactants include surfactants that areblends of sucrose fatty acid esters, containing a plurality of differentsucrose fatty acid esters. The different sucrose fatty acid esters inthe blend can vary in the length and/or saturation of the carbon chainof the fatty acid portion of the ester, or in the degree ofesterification (e.g., whether the ester is a monoester, diester,triester, or polyester). Typically, the sucrose fatty acid estersurfactant contains proportionally more monoesters than other types ofesters (e.g., diesters, triesters, and polyesters).

The relative amount of monoester in the sucrose fatty acid estersurfactant can be specified as a percentage of the total esters. Forexample, among the sucrose fatty acid ester surfactants are surfactantscontaining at least at or about 50%, at least at or about 60%, at leastat or about 70%, at least at or about 80%, or at least at or about 90%,by weight or by molecule, sucrose fatty acid monoesters. In one aspect,the sucrose fatty acid ester surfactant is a blend of sucrose fatty acidesters containing at or about 72% monoesters, at or about 23% diesters,and at or about 5% triesters, by weight or by molecule. In anotheraspect, the sucrose fatty acid ester surfactant is a blend of sucrosefatty acid esters containing at or about 61%, monoesters, at or about30% diesters, at or about 7%, triesters, and at or about 2%, polyesters,by weight or by molecule. In another aspect, the sucrose fatty acidester surfactant is a blend of sucrose fatty acid esters containing ator about 52%, monoesters, at or about 36%, diesters, at or about 10%triesters, and at or about 2% polyesters, by weight or by molecule.

The sucrose fatty acid ester(s) of the sucrose fatty acid estersurfactants can contain a fatty acid chain of any length, and typicallyhave between 4 or about 4 and 28 or about 28 carbon atoms, typicallybetween 8 or about 8 and 22 or about 22 carbon atoms (e.g., 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 carbon atoms), andmore typically between 12 or about 12 and 18 or about 18 carbon atoms(e.g., 12, 13, 14, 15, 16, 17, or 18 carbon atoms), such as fatty acidshaving carbon chains that contain 12, 14, 16 or 18 carbon atoms, such asstearic acid, lauric acid, oleic acid, and palmitic acid. Such sucrosefatty acid esters include sucrose stearate (e.g., sucrose monostearate),sucrose laurate (e.g., sucrose monolaurate), sucrose oleate (e.g.,sucrose monooleate), sucrose palmitate (e.g., sucrose monopalmitate),and combinations thereof, including homologs thereof. The sucrose fattyacid ester surfactants include any of those described herein, andtypically those having an HLB value of between 14 or about 14 and 20 orabout 20, and more typically between 15 or about 15 and 18 or about 18(e.g., at or about 15, 16, 17, or 18). In one example, the sucrose fattyacid ester surfactant contains any one or more of sucrose monostearate,sucrose monolaurate, sucrose monooleate, and sucrose monopalmitate.Exemplary sucrose fatty acid esters are described herein, and includethose having a structure according to Scheme V, provided herein below.

Exemplary non-polar compounds in the provided compositions (includingthe liquid nanoemulsion concentrates) are non-polar active ingredients,which include, but are not limited to, omega-3 fatty acids, omega-6fatty acids, conjugated fatty acids, Coenzyme Q10 (e.g., ubidecarenone),phytosterols, and saw palmetto extracts. The non-polar activeingredients include, for example, non-polar compounds containingDocosahexaenoic acid (DHA) and/or Eicosapentaenoic acid (EPA),Alpha-Linolenic acid (α-Linolenic acid; ALA), conjugated linoleic acid(CLA), and gamma-linolenic acid (GLA), including, but not limited to,fish oil, algae oil, flaxseed oil, borage oil, and saw palmetto extract.

The non-polar active ingredients include, but are not limited to,compounds containing any fat-soluble nutraceutical or pharmaceuticaland/or oil, such as, for example, drugs, hormones, vitamins, nutrients,including any and other lipophilic compounds containing essential fattyacids, for example, polyunsaturated fatty acids (PUFAs), including, forexample, omega-3 fatty acids, for example, natural and synthetic omega-3fatty acids, for example, compounds containing omega-3 polyunsaturatedlong-chain fatty acids, including Eicosapentaenoic acid (EPA) (20:5ω3),Docosahexaenoic acid (DHA) (22:6ω3), Eicosatetraenoic acid (24:4ω3);Docosapentaenoic acid (DPA, Clupanodonic acid) (22:5ω3), 16:3 ω3; 24:5ω3, and/or nisinic acid (24:6ω3), which can include, for example, fishoil, algae oil, krill oil, canola oil, flaxseed oil, soybean oil, andwalnut oil; compounds containing short-chain omega-3 fatty acids, forexample, Alpha-Linolenic acid (α-Linolenic acid; ALA) (18:3ω3) (e.g.,flaxseed oil) and Stearidonic acid (18:4ω3); esters of an omega-3 fattyacid and glycerol, for example, monoglycerides, diglycerides andtriglycerides; esters of omega-3 fatty acid and a primary alcohol, forexample, fatty acid methyl esters and fatty acid esters; precursors ofomega-3 fatty acid oils, for example, EPA precursor, DHA precursor;derivatives such as polyglycolized derivatives or polyoxyethylenederivatives; oils containing the omega-3 fatty acids, for example, fishoil (e.g., marine oil), including, for example, highly purified fish oilconcentrates, perilla oil, krill oil, and algae oil (e.g., microalgaeoil); compounds containing omega-6 fatty acids, for example, compoundscontaining Linoleic acid (18:2ω6) (a short-chain fatty acid),Gamma-linolenic acid (GLA) (18:3ω6), Dihomo gamma linolenic acid (DGLA)(20:3ω6), Eicosadienoic acid (20:2ω6), Arachidonic acid (AA) (20:4ω6),Docosadienoic acid (22:2ω6), Adrenic acid (22:4ω6), and/orDocosapentaenoic acid (22:5ω6), for example, borage oil, corn oil,cottonseed oil, grape seed oil, peanut oil, primrose oil, for example,evening primrose (Oenothera biennis) oil, blackcurrant seed oil, hempseed oil, spirulina extract, safflower oil, sesame oil and soybean oil;

compounds containing other fatty acids, for example, triglycerides,including medium chain triglycerides, polar lipids, for example, etherlipids, phosphoric acid, choline, fatty acids, glycerol, glycolipids,triglycerides, and phospholipids (e.g., phosphatidylcholine (lecithin),phosphatidylethanolamine, and phosphatidylinositol); saw palmettoextract; and ethyl linoleate; and herb oils, for example, garlic oilsand scordinin; short-chain saturated fatty acids (4:0-10:0), Lauric acid(12:0), Myristic acid (14:0), Pentadecanoic acid (15:0), Palmitic acid(16:0), Palmitoleic acid (16:1 ω7), Heptadecanoic acid (17:0), Stearicacid (18:0), Oleic acid (18:1 ω9), Arachidic acid (20:0);

compounds containing micronutrients, for example, vitamins, minerals,co-factors, for example, coenzymes, such as coenzyme Q, e.g., CoenzymeQ10 (CoQ10, also called ubiquinone, e.g., ubidecarenone or a reducedform of CoQ10, e.g., ubiquinol), tumeric extract (e.g., cucuminoids),saw palmetto lipid extract (e.g., saw palmetto oil) echinacea extract,hawthorn berry extract, ginseng extract, lipoic acid (e.g., thiocticacid), ascorbyl palmitate, kava extract, St. John's Wort (e.g.,hypericum, Klamath weed, goat weed), extract of quercitin,dehydroepiandrosterone, indol-3-carbinol;

compounds containing carotenoids, including hydrocarbons and oxygenated,alcoholic derivatives of hydrocarbons, for example, beta carotene, mixedcarotenoids complex, lutein, lycopene, Zeaxanthin, Cryptoxanthin, forexample, beta-crytoxanthin, beta carotene, mixed carotenoids complex,astaxanthin, bixin, canthaxanthin, capsanthin, capsorubin,apo-carotenal, beta-12′-apo-carotenal, “Carotene” (mixture of alpha andbeta-carotene), gamma carotene, violerythrin, zeaxanthin, esters ofhydroxyl- or carboxyl-containing members thereof;

compounds containing fat-soluble vitamins, for example, Vitamins A, D, Eand K, and corresponding provitamins and vitamin derivatives such asesters with an action resembling that of vitamin A, D, E or K forexample; retinol (vitamin A) and pharmaceutically acceptable derivativesthereof, for example, palmitate ester of retinol and other esters ofretinol, and calciferol (vitamin D, including vitamin D3 ergocalciferoland/or vitamin D3 cholecalciferol) and its pharmaceutically acceptablederivatives thereof and precursors of vitamin D, d-alpha tocopherol(vitamin E) and derivatives thereof, including pharmaceuticalderivatives thereof, for example, Tocotrienols, d-alpha tocopherolacetate and other esters of d-alpha tocopherol, and ascorbyl palmitate,a fat-soluble version of vitamin C;

compounds containing phytochemicals, including phytoestrogens, forexample, genistein and daidzein, for example, isoflavones, for example,soy isoflavones, flavonoids, phytoalexins, for example, Resveratrol(3,5,4′-trihydroxystilbene), red clover extract, and phytosterols;

compounds containing lipid-soluble drugs, including natural andsynthetic forms of immunosuppressive drugs, such as Cyclosporin,protease inhibitors such as Ritonavir, macrolide antibiotics and oilsoluble anesthetics such as Propofol, natural and synthetic forms ofsteroidal hormones, for example, estrogens, estradiols, progesterone,testosterone, cortisone, phytoestrogens, dehydroepiandrosterone (DHEA),growth hormones and other hormones;

compounds containing oil-soluble acids and alcohols, for example,tartaric acid, lactylic acid butylated hydroxyanisole, butylatedhydroxytoluene, lignin, sterols, polyphenolic compounds, oryzanol,cholesterol, phytosterols, flavonoids, such as, but not limited to,quercetin and reservatol, and diallyl disulfides.

The non-polar active ingredients include ingredients containingpolyunsaturated fatty acids, such as compounds containing any one ormore of omega-3 fatty acids, including docosahexaenoic acid (DHA),eicosapentaenoic acid (EPA), and alpha-linolenic acid (ALA) (e.g., fishoils, krill oils, algae oils, and/or flaxseed oils), omega-6 fattyacids, such as gamma-linolenic acid (GLA) (e.g., borage oils);conjugated fatty acids (e.g., conjugated linoleic acid (CLA)), and sawpalmetto extracts; and ingredients containing coenzymes such as coenzymeQ, for example, Coenzyme Q10 (e.g., ubidecarenone); and ingredientscontaining phytosterols, and combinations thereof.

In one example, the non-polar active ingredient containseicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or acombination thereof. In one example, the non-polar active ingredientcontains DHA at an amount between 20% or about 20% and 90% or about 90%;between 25% or about 25% and 85% or about 85%; between 35% or about 35%and 70% or about 70%; or between 25% or about 25% and 40% or about 40%,by weight (w/w), of the non-polar active ingredient. In another aspect,the non-polar active ingredient contains EPA at an amount between 5% orabout 5% and 15% or about 15%; between 5% or about 5% and 13% or about13%; or between 5% or about 5% and 10% or about 10% by weight (w/w), ofthe non-polar active ingredient. In one aspect, the amount of EPA is notmore than 10% or about 10%, or not more than 13% or about 13%, by weight(w/w), of the non-polar active ingredient. For example, the non-polaractive ingredients include fish oil and algae oil containing any suchpercentage of EPA and/or DHA.

In another example, the non-polar active ingredient containsalpha-linolenic acid (ALA). In one example, the non-polar activeingredient contains ALA at an amount of at least 50% or about 50%, byweight (w/w), of the non-polar active ingredient, such as between 50% orabout 50% and 80% or about 80%, or between 65% or about 65% and 75% orabout 75%, by weight (w/w), of the non-polar active ingredient. Forexample, the non-polar active ingredients include flaxseed oilscontaining any such percentage of ALA.

In another example, the non-polar active ingredient containsgamma-linolenic acid (GLA). In one example, the non-polar activeingredient contains GLA at an amount of at least 22% or about 22%, byweight (w/w), of the non-polar active ingredient. For example, thenon-polar active ingredients include a borage oil containing GLA at anamount of at least 22% or about 22%, by weight (w/w), of the borage oil.

In some examples, the concentrate contains more than one non-polaractive ingredient, for example, two or more non-polar activeingredients, such as any of the non-polar compounds described herein. Inone example, the total amount of non-polar active ingredient(s) isbetween at or about 5% and 10% of the weight of the concentrate, forexample, where the combined weight of the non-polar active ingredientand additional non-polar active ingredient(s) is not more than at orabout 10%, by weight (w/w), of the concentrate.

The polar solvents contained in the concentrates include polar proticand polar aprotic solvents, and typically are polar protic solvents,such as polar solvents having a dielectric constant greater than 15 orabout 15 or equal to 15 or about 15, or a dielectric constant between 20or about 20 and 90 or about 90, such as between 20 or about 20 and 80 orabout 80 (e.g., at or about or at least at or about 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, or 81); and polar solvents having a polarity index of between ator about 3 and at or about 9 or a dipole moment between at or about 1.8and at or about 2.8. The polar solvents include water and alcohols, suchas monohydric, dihydric, trihydric and other alcohols, and typicallyalcohols other than monohydric alcohols, alcohols having two or morehydroxyl groups, such as dihydric (two hydroxy groups) and trihydric(three hydroxyl groups) alcohols. The polar solvents include, but arenot limited to, glycerin, ethylene glycols, such as propylene glycol,ethylene glycol, tetraethylene glycol, triethylene glycol, andtrimethylene glycol. Polar solvents can further include low molecularweight polyethylene glycols (PEGs), such as PEGs with molecular weightsat or about, or less than at or about, 600, 400 or 200 kDa. In someexamples, the polar solvent is water, glycerin, or propylene glycol.

The provided concentrates can contain one or more additionalingredients. In one example, the concentrate further contains aco-surfactant in an amount sufficient to stabilize the concentrate,compared to the absence of the co-surfactant. In one aspect, theco-surfactant is a phospholipid, such as, but not limited to, aphosphatidylcholine. In one example, the amount of the co-surfactant,e.g., the phospholipid, is between 0.1% or about 0.1% and 1% or about1%, by weight (w/w), of the concentrate.

In another example, the concentrate further contains a preservative, inamount sufficient to preserve the concentrate, compared to the absenceof the preservative. Exemplary preservatives in the compositionsprovided are natural preservatives, such as benzyl alcohol andpreservatives containing benzyl alcohol. In one example, the amount ofpreservative is between 0.1% or about 0.1% and 1% or about 1%, by weight(w/w), of the concentrate, for example, at or about 0.1, 0.2, 0.3, 0.4,0.5, 0.6, 0.7, 0.8, 0.9, or 1%, by weight of the concentrate. In oneexample, the amount of benzyl alcohol is between 0.1% or about 0.1% and1% or about 1%, by weight (w/w), of the concentrate, for example, at orabout 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1%, by weight ofthe concentrate.

In another example, the concentrate contains a non-polar solvent, forexample, a non-polar solvent that dissolves the non-polar activeingredient and differs therefrom. Typically, the amount of non-polarsolvent is sufficient to dissolve the non-polar active ingredient, andcan be, for example, between 1% or about 1% and 6% or about 6%, forexample, at or about 1, 2, 3, 4, 5, or 6%, by weight (w/w), of theconcentrate. The non-polar solvent typically is an oil, such as any oilsuitable for dissolving the non-polar ingredient. Exemplary non-polarsolvents in the compositions provided are Vitamin E oil, flaxseed oil,sunflower oil, any vegetable oils, and other oils.

In another example, the concentrate contains an emulsion stabilizer.Typically, the emulsion stabilizer is included in the concentrate at anamount sufficient to stabilize the concentrate. The emulsion stabilizersinclude, but are not limited to, compositions containing a blend ofgums, such as the Saladizer® brand emulsion stabilizer. In one example,the emulsion stabilizer contains one or more of guar gum, xanthan gumand sodium alginate. In one example, the emulsion stabilizer containsguar gum, xanthan gum, and sodium alginate.

In another example, the concentrates contain flavors. Typically, theflavor(s) is/are included at an amount sufficient to enhance the tasteof the concentrate, the smell of the concentrate, or a combinationthereof, compared to the absence of the flavor. The flavors include, butare not limited to, flavors containing lemon oil and D-limonene, orcombination thereof, or any other known flavors, such as flavorsdescribed herein.

In another example, when the concentrates contains water as the polarsolvent, for example, the concentrates contain pH adjusters. Typically,the pH adjuster contains an acid or a base at an amount sufficient toaffect the pH of the concentrate compared to the absence of the pHadjuster. The pH adjusters include, but are not limited to, citric acidand phosphoric acid.

The provided concentrates include, but are not limited to, concentratescontaining PUFA-containing non-polar active ingredients, such as omega-3fatty acid, omega-6 fatty acid, conjugated fatty acid, and saw palmettooil containing non-polar active ingredients.

In one particular example, the provided concentrate contains a non-polaractive ingredient that is a fish oil containing EPA and DHA (e.g., afish oil containing 10% EPA and 70% DHA; a fish oil containing about 13%and about 13% DHA; or a fish oil containing 40% EPA and 20% DHA), asurfactant that is a sucrose fatty acid ester (SFAE) surfactantcontaining monoesters (e.g., that sold under the name DK Ester® F-160 orF-140 SFAE, produced by Dai-Ichi Kogyo Seiyaku Co., Ltd of Japan anddistributed through Montello Inc., Tulsa, Okla.), and a polar solventthat is water. In one aspect of this example, the amount of the fish oilnon-polar active ingredient is at or about 5%, by weight (w/w), of theconcentrate, the amount of water is at or about 69.02%, by weight (w/w),of the concentrate, and the amount of the SFAE surfactant is at or about25.2%, by weight (w/w), of the concentrate. In one aspect, a portion ofthe amount of the SFAE is added to the water phase and a portion of theamount of the SFAE surfactant is added to the oil phase, for example,22.7%, by weight (w/w), SFAE in the water phase and 2.5%, by weight inthe oil phase. In one aspect of this example, the concentrate furthercontains a benzyl alcohol preservative and a pH adjuster that is citricacid.

In another example, the concentrate contains a non-polar activeingredient that is a flaxseed oil (e.g., one containing 50% or 55% omega3 fatty acids, e.g., 50% or 55% ALA), a surfactant that is a sucrosefatty acid ester (SFAE) surfactant containing monoesters (e.g., thatsold under the name DK Ester® F-160 or F-140 SFAE, produced by Dai-IchiKogyo Seiyaku Co., Ltd of Japan and distributed through Montello Inc.,Tulsa, Okla.), and a polar solvent that is water. In one aspect of thisexample, the amount of the flaxseed oil non-polar active ingredient isat or about 5%, by weight (w/w), of the concentrate, the amount of wateris at or about 69.02%, by weight (w/w), of the concentrate, and theamount of the SFAE surfactant is at or about 25.2%, by weight (w/w), ofthe concentrate. In one aspect, a portion of the amount of the SFAE isadded to the water phase and a portion of the amount of the SFAEsurfactant is added to the oil phase, for example, 22.7%, by weight(w/w), SFAE in the water phase and 2.5%, by weight in the oil phase. Inone aspect, the concentrate further contains a benzyl alcoholpreservative, and a pH adjuster that is citric acid.

In another example, the concentrate contains a non-polar activeingredient that contains gamma linolenic acid (GLA) (e.g., a borage oilcontaining GLA, e.g., at 22%), a surfactant that is a sucrose fatty acidester (SFAE) surfactant containing monoesters (e.g., that sold under thename DK Ester® F-160 or F-140 SFAE, produced by Dai-Ichi Kogyo SeiyakuCo., Ltd of Japan and distributed through Montello Inc., Tulsa, Okla.),and a polar solvent that is water. In one aspect of this example, theamount of the GLA-containing non-polar active ingredient is at or about5%, by weight (w/w), of the concentrate, the amount of water is at orabout 69.02%, by weight (w/w), of the concentrate, and the amount of theSFAE surfactant is at or about 25.2%, by weight (w/w), of theconcentrate. In one aspect, a portion of the amount of the SFAE is addedto the water phase and a portion of the amount of the SFAE surfactant isadded to the oil phase, for example, 22.7%, by weight (w/w), SFAE in thewater phase and 2.5%, by weight in the oil phase. In one aspect, theconcentrate further contains a benzyl alcohol preservative.

In another example, the concentrate contains a non-polar activeingredient that conjugated linoleic acid (CLA) (e.g., 80% CLA), asurfactant that is a sucrose fatty acid ester (SFAE) surfactantcontaining monoesters (e.g., that sold under the name DK Ester® F-160 orF-140 SFAE, produced by Dai-Ichi Kogyo Seiyaku Co., Ltd of Japan anddistributed through Montello Inc., Tulsa, Okla.), and a polar solventthat is water. In one aspect of this example, the amount of theCLA-containing non-polar active ingredient is at or about 5%, by weight(w/w), of the concentrate, the amount of water is at or about 69.02%, byweight (w/w), of the concentrate, and the amount of the SFAE surfactantis at or about 25.2%, by weight (w/w), of the concentrate. In oneaspect, a portion of the amount of the SFAE is added to the water phaseand a portion of the amount of the SFAE surfactant is added to the oilphase, for example, 22.7%, by weight (w/w), SFAE in the water phase and2.5%, by weight in the oil phase. In one aspect of this example, theconcentrate further contains a benzyl alcohol preservative, and a pHadjuster that is citric acid.

In another example, the concentrate contains a CoQ10 non-polar activeingredient, a surfactant that is a sucrose fatty acid ester (SFAE)surfactant containing monoesters (e.g., that sold under the name DKEster® F-160 or F-140 SFAE, produced by Dai-Ichi Kogyo Seiyaku Co., Ltdof Japan and distributed through Montello Inc., Tulsa, Okla.), and apolar solvent that is water. In one aspect of this example, the amountof the CoQ10 non-polar active ingredient is at or about 5%, by weight(w/w), of the concentrate, the amount of water is at or about 69.02%, byweight (w/w), of the concentrate, and the amount of the SFAE surfactantis at or about 25.2%, by weight (w/w), of the concentrate. In oneaspect, a portion of the amount of the SFAE is added to the water phaseand a portion of the amount of the SFAE surfactant is added to the oilphase, for example, 22.7%, by weight (w/w), SFAE in the water phase and2.5%, by weight (w/w), in the oil phase. In one aspect, the concentratefurther contains a benzyl alcohol preservative. In another aspect, theconcentrate further contains D-limonene and lemon oil flavors. Inanother aspect, the concentrate further contains a phosphatidylcholineco-surfactant. In another aspect, the concentrate further contains anon-polar solvent that is a Vitamin E oil or other oil. In anotherexample, the concentrate further contains an emulsion stabilizercontaining a blend of gums, such as a blend of xanthan gum, guar gum,and/or sodium alginate.

In some examples, the concentrate is formulated based on the desiredproperties of resulting dilution compositions generated by diluting theconcentrate in an aqueous liquid. Typically, the concentrate is formedso that it can be diluted in aqueous medium to produce a liquid dilutioncomposition having one, more than one, all, or any combination of, ofthe following properties:

In one example, the concentrate is formulated such that dilution of acertain amount of the concentrate in a certain amount of aqueous mediumyields a liquid dilution composition having a desired particle size,typically a particle size that is not greater than a particular particlesize or is less than a particular particle size. The specified particlesize can be expressed as the average particle size, or the largestparticle size in the aqueous medium. For example, it can be desired thatthe liquid dilution composition contains less than a particular particlesize on average or at most. For example, the concentrate can beformulated such that dilution of at least 0.5 grams (g) or about 0.5 g,at least 1 g or about 1 g, at least 2 g or about 2 g, at least 5 g orabout 5 g, or at least 10 g or about 10 g of the concentrate into at orabout 8 fluid ounces (0.236588 liters) of an aqueous medium; or dilutionof the concentrate in an aqueous medium, at a dilution not more than1:10 or about 1:10, not more than 1:25 or about 1:25, not more than 1:50or about 1:50, not more than 1:100 or about 1:100, not more than 1:250or about 1:250 or, at most, not more than 1:500 or about 1:500; ordilution of the concentrate in an aqueous medium to form a liquiddilution composition containing at least 25 mg or about 25 mg, at least35 mg or about 35 mg, at least 50 mg or about 50 mg, at least 100 mg orabout 100 mg, at least 250 mg or about 250 mg, or at least 500 mg orabout 500 mg of the non-polar active ingredient per 8 fluid ounces ofthe liquid dilution composition, yields a liquid dilution compositionhaving a particle size of less than 500 nm or less than about 500 nm,less than 200 nm or less than about 200 nm, less than 100 nm or lessthan about 100 nm, less than 50 nm or less than about 50 nm or less than25 nm or less than about 25 nm, at most or on average.

In another example, the concentrate is formulated such that dilution ofa certain amount of the concentrate in an amount of aqueous mediumyields a liquid dilution composition having a desired clarity, such asby yielding a dilution composition having a desired NTU value, typicallyan NTU value that is not greater than or is less than a given NTU value,or by yielding a liquid dilution composition that is as clear or aboutas clear as the aqueous medium prior to the addition of the concentrate(i.e., in the absence of the concentrate). For example, the concentratecan be formulated such that dilution of at least 0.5 grams (g) or about0.5 g, at least 1 g or about 1 g, at least 2 g or about 2 g, at least 5g or about 5 g, or at least 10 g or about 10 g of the concentrate intoat or about 8 fluid ounces (0.236588 liters) of an aqueous medium; ordilution of the concentrate in an aqueous medium, at a dilution not morethan 1:10 or about 1:10, not more than 1:25 or about 1:25, not more than1:50 or about 1:50, not more than 1:100 or about 1:100, not more than1:250 or about 1:250 or, at most, not more than 1:500 or about 1:500; ordilution of the concentrate in an aqueous medium to form a liquiddilution composition containing at least 25 mg or about 25 mg, at least35 mg or about 35 mg, at least 50 mg or about 50 mg, at least 100 mg orabout 100 mg, at least 250 mg or about 250 mg, or at least 500 mg orabout 500 mg of the non-polar active ingredient per 8 fluid ounces ofthe liquid dilution composition, yields a liquid dilution compositionhaving a Nephelometric Turbidity Units (NTU) value of less than 200 orabout 200, less than 100 or about 100, less than 50 or about 50, lessthan 30 or about 30, less than 25 or about 25, or less than 10 or about10, or yields a liquid dilution composition that is at least as clear orat least about as clear as, the aqueous medium in the absence of theconcentrate (i.e., compared to the clarity of the aqueous medium priorto addition of the concentrate).

In another example, the concentrate is formulated such that, upondilution, it yields a stable liquid dilution composition, for example acomposition that does not contain visible particles, does not containvisible crystals, does not exhibit ringing, or a combination thereof.The stability can be for a specified period of time, and/or when theconcentrate or liquid dilution composition is kept at a particulartemperature. For example, the concentrate can be formulated such thatdilution of at least 0.5 g or about 0.5 g, at least 1 g or about 1 g, atleast 2 g or about 2 g, at least 5 g or about 5 g, or at least 10 g orabout 10 g of the concentrate into 8 fluid ounces (0.236588 liters), orabout 8 fluid ounces, of an aqueous medium; dilution of the concentratein an aqueous medium, at a dilution of not more than 1:10 or about 1:10,not more than 1:25 or about 1:25, not more than 1:50 or about 1:50, notmore than 1:100 or about 1:100, not more than 1:250 or about 1:250, or,at most, not more than 1:500 or about 1:500; and/or dilution of theconcentrate into an aqueous medium to form a liquid dilution compositioncontaining at least 25 mg or about 25 mg, at least 35 mg or about 35 mg,at least 50 mg or about 50 mg, at least 100 mg or about 100 mg, at least250 mg or about 250 mg, or at least 500 mg or about 500 mg of thenon-polar active ingredient per 8 fluid ounces of the liquid dilutioncomposition, yields a liquid dilution composition that does not containvisible particles, does not contain visible crystals, does not exhibitphase separation, and/or does not exhibit ringing, and/or is pleasanttasting and/or smelling. The concentrate can be formulated such that theliquid dilution composition remains free from visible particles, remainsfree from visible crystals, remains free from phase separation, remainsfree from ringing and/or is pleasant tasting and smelling, when theconcentrate and/or the liquid dilution composition is stored at roomtemperature, or at a refrigerated temperature, or at a frozentemperature. The storage can be, for example, for at least one day, atleast one week, at least thirty days, or at least one year.

The aqueous medium can be a beverage, such as, for example, water,juice, soda, tea, coffee, sports drinks, nutritional beverages, energydrinks, milk, and other beverages, including those described herein.

Also provided are the liquid dilution compositions, which contain theconcentrates diluted in an aqueous medium, e.g., a beverage. The liquiddilution compositions can contain any of the provided concentrates.Thus, the liquid dilution compositions contain the non-polar activeingredients in aqueous medium, such as beverages, that are desirable forhuman consumption. The liquid dilution compositions include those madeby diluting the concentrates, such as those having the properties asdescribed above, such as the desired particle size, clarity, NTU valueand/or stability, for example, lack of ringing, visible crystals, phaseseparation, and/or pleasant taste/smell, for example, according to thespecifications described above.

In one example, the aqueous medium contained in the liquid dilutioncomposition is a beverage, such as, for example, water, soda, milk, tea,coffee, juice, energy drink or a sports or nutrition beverage. In oneaspect, the liquid dilution composition is as clear or about as clear asthe aqueous medium, such as the beverage, prior to addition of theconcentrate (e.g., compared to the absence of the concentrate), and/orremains as clear or about as clear as the beverage when stored at roomtemperature (e.g., 25° C. or about 25° C.), or at a refrigeratedtemperature (e.g., 0-10° C. or about 0-10° C., e.g., at or about 4° C.),or at a frozen temperature (e.g., −20° C. or about −20° C.), wherein thestorage is for at least one day, at least one week, at least thirtydays, or at least one year.

The amount of the concentrate in the liquid dilution composition can bespecified. For example, the liquid dilution compositions include thosecontaining at least 0.5 grams (g) or about 0.5 g, at least 1 g or about1 g, at least 2 g or about 2 g, at least 5 g or about 5 g, or at least10 g or about 10 g of the concentrate, per 8 fluid ounces (0.236588liters) of the aqueous medium; or containing the concentrate at adilution of not more than 1:10 or about 1:10, not more than 1:25 orabout 1:25, not more than 1:50 or about 1:50, not more than 1:100 orabout 1:100, not more than 1:250 or about 1:250 or, at most, not morethan 1:500 or about 1:500; or containing at least 25 mg or about 25 mg,at least 35 mg or about 35 mg, at least 50 mg or about 50 mg, at least100 mg or about 100 mg, at least 250 mg or about 250 mg, or at least 500mg or about 500 mg of the non-polar active ingredient per 8 fluid ouncesof the aqueous medium. The liquid dilution compositions typically haveone or more desired property, such as particle size, clarity, NTU value,stability, e.g., free from crystals, phase separation, ringing orunpleasant taste/smell, such as for at least a specified amount of timewhen stored under specified storage conditions.

For example, the compositions include liquid dilution compositionshaving a particle size less than 500 or about 500, less than 300 orabout 300, less than 200 or about 200 nm, less than 100 or about 100 nm,less than 50 or about 50 nm or less than 25 or about 25 nm on theaverage or at the most; those having an NTU value less than 500 or about500, less than 300 or about 300, less than 200 or about 200, less than100 or about 100, less than 50 or about 50, less than 25 or about 25, orless than 10 or about 10; those containing visible particles, does notcontain visible crystals, not exhibiting ringing and/or phaseseparation; and/or remaining free from (or does not exhibit) visibleparticles, visible crystals, ringing and/or phase separation, and/orunpleasant taste/smell when stored at room temperature (e.g., 25° C. orabout 25° C.), or at a refrigerated temperature (e.g., 0-10° C. or about0-10° C., e.g., at or about 4° C.), or at a frozen temperature (e.g.,−20° C. or about −20° C.), when the storage is for at least one day, atleast one week, at least thirty days, or at least one year.

Also provided are methods for making the concentrates and methods formaking the liquid dilution compositions. Generally, the methods formaking the concentrates are carried out by generating, separately, anoil phase and a water phase, and mixing the two phases, typically byemulsification, to form the concentrate, which is a liquid nanoemulsionconcentrate. Oil phase ingredients are added to form the oil phase andwater phase ingredients are added to form the water phase. Theingredients are selected from ingredients of the concentrates, asdescribed herein, which typically include a non-polar compound, asurfactant and a polar solvent, as described herein. Typically, the oilphase ingredients include the non-polar compound(s), typically anon-polar active ingredient(s), of the concentrate, and the water phaseingredients include the polar solvent. The ingredients are added atamounts within the appropriate concentration range for the providedconcentrates as described herein. In one example, the water phaseingredients include the surfactant. In another example, the oil phaseingredients contain the surfactant. In one example, the water phaseingredients and the oil phase ingredients contain the surfactant.

The amounts of the surfactant(s), non-polar active ingredient(s) andpolar solvent are selected based on the appropriate concentration rangesof these ingredients in the resulting concentrate. For example, thenon-polar active ingredient is included at an amount that is between 5%or about 5% and 10% or about 10%, by weight (w/w), of the finalconcentrate; the surfactant is included at an amount that is between 16%or about 16% and 30% or about 30%, by weight (w/w), of the finalconcentrate; and the polar solvent is included at an amount that isbetween 60% or about 60% and 79% or about 79%, by weight (w/w), of thefinal concentrate, as described above.

In one example, the oil phase ingredients further include the non-polarsolvent(s). In one example, the concentrate is made with first andsecond oil phase ingredients and the first oil phase ingredients includethe non-polar active ingredient and the solvent. In one example, thesolvent contains an oil, other than the non-polar active ingredient,such as, for example, Vitamin E, flaxseed oil and/or safflower oil.

In one example, the oil phase ingredients and/or the water phaseingredients contain the co-surfactant, at an amount sufficient tostabilize the concentrate, such as a phospholipid, e.g.,phosphatidylcholine. In one example, the amount of phospholipid isbetween 0.1% or about 0.1% and 1% or about 1%, by weight (w/w), of theconcentrate. In another example, the oil phase ingredients and/or thewater phase ingredients further contain the at least one preservative inamount sufficient to preserve the concentrate, such as, for example, apreservative containing benzyl alcohol. In one example, the amount ofpreservative and/or the benzyl alcohol is between 0.1% or about 0.1% and1% or about 1%, by weight (w/w), of the concentrate.

In another example, the oil phase ingredients and/or the water phaseingredients further contain an emulsion stabilizer, at an amountsufficient to stabilize the concentrate, such as an emulsion stabilizercontaining a blend of gums, such as any one or more of guar gum, xanthangum and sodium alginate.

In an exemplary provided method for making the concentrate, an oil phaseis generated by mixing the oil phase ingredients in a first vessel andheating the oil phase ingredients; a water phase is generated by mixingone or more water phase ingredients in a second vessel and heating thewater phase ingredients; and the oil and water phases are emulsified togenerate the concentrate.

In another exemplary provided method, an oil phase is generated bymixing one or more first oil phase ingredients in a first vessel andheating the first oil phase ingredients at least until the first oilphase ingredients dissolve; then adding one or more additional oil phaseingredients to the first vessel; and mixing and heating the first andthe additional oil phase ingredients; a water phase is generated bymixing one or more water phase ingredients in a second vessel andheating the water phase ingredient(s); and the water and oil phases areemulsified, to generate the concentrate.

The heating and mixing of the water and oil phases can be carried outsimultaneously or sequentially, in any order.

In any of the provided methods for making the concentrates, the mixingsteps (e.g., mixing the oil and/or water phases) can be carried out witha standard mixer, such as any of the standard mixers listed herein, orwith any of the other mixers described herein, such as with ahomogenizer. In any of the provided methods, the heating can be carriedout using one or more heating apparatuses, such as, for example, a hotplate, a water jacket, or any of the heating apparatuses listed herein.In one example, the oil phase ingredients are heated with a firstheating apparatus and the water phase ingredients are heated with asecond heating apparatus. In one example, heating involves heating theingredients to 60° C. or about 60° C., or to at or about 70° C. or at orabout 71° C. In one example, the oil phase and/or water phaseingredients are heated to between about between 45° C. or about 45° C.and 85° C. or about 85° C., for example, at or about 45, 46, 47, 48, 49,50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or85° C.

In any of the provided methods, the emulsifying can be carried out usinga homogenizer, such as any homogenizer described herein. In one example,the emulsifying is performed at between 850 rpm or about 850 rpm and1200 rpm or about 1200 rpm. In another example, the emulsifying isperformed at a speed lower then 850 rpm, such as, for example, between25 or about 25 rpm and 50 rpm or about 50 rpm, for example at or about30 rpm.

In some examples, the methods further include rapidly cooling theforming emulsion during the emulsifying step. In some examples, rapidcooling results in cooling of the forming emulsion to between 25° C. orabout 25° C. and 43° C. or about 43° C., such as 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42 or 43° C., e.g., betweenat or about 25° C. and at or about 35° C., or between at or about 35° C.and at or about 43° C. In one example, the cooling of the emulsionresults in less than at or about 60 minutes, in less than at or about 30minutes, or between at or about 30 and at or about 60 minutes. Exemplaryof the means for performing the rapid cooling include repeatedly passingthe phases through a cooling apparatus attached to a vessel.

The provided methods further can include adding one or more flavors(e.g., lemon oil, D-limonene) and/or one or more pH adjusters (e.g.,citric acid, phosphoric acid) to the concentrate, for example, afteremulsifying the oil and water phases. The pH typically is measuredsimultaneously with, before and/or after addition of the pH adjuster,and the amount of pH adjuster is determined by the pH of theconcentrate. Typically, the pH adjuster comprises an acid or a base atan amount sufficient to affect the pH of the concentrate.

In one example of the methods, the ingredients are added to thevessel(s) simultaneously or sequentially, in any order. In anotherexample, the ingredients (e.g., the oil phase and/or water phaseingredients) are added to the vessels in a particular order, such as aspecific order provided herein, for example, in the individual examplesprovided. In one example, where the water phase ingredients contain apolar solvent (e.g., water, propylene glycol, glycerin or other diolsuch as another sugar alcohol) and an emulsion stabilizer, the waterphase ingredients are added sequentially, in the following order: 1)polar solvent (e.g., water, propylene glycol or glycerin); 2) emulsionstabilizer. In another example, where the oil phase ingredients containthe surfactant, the non-polar compound and a preservative, the oil phaseingredients are added sequentially, in the following order: 1)surfactant, 2) preservative; 3) non-polar compound (i.e., non-polaractive ingredient). In another example, where the oil phase ingredientscontain the surfactant, the non-polar compound, a preservative and anemulsion stabilizer, the oil phase ingredients are added to the oilphase vessel sequentially, in the following order: 1) surfactant; 2)preservative; 3) non-polar compound; and 4) emulsion stabilizer. Inanother example, where the oil phase ingredients contain the surfactant,the non-polar compound, a preservative, a solvent and an emulsionstabilizer, the oil phase ingredients are added to the oil phase vesselsequentially, in the following order: 1) surfactant; 2) preservative; 3)non-polar solvent; 4) non-polar compound; and 5) emulsion stabilizer. Inanother example, where the oil phase ingredients contain the surfactant,the non-polar compound, a preservative, a non-polar solvent, aco-surfactant and an emulsion stabilizer, the oil phase ingredients areadded to the oil phase vessel sequentially, in the following order: 1)surfactant; 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5)non-polar compound; and 6) emulsion stabilizer. In another example,where the water phase contains a polar solvent and a surfactant, thepolar solvent and the surfactant are added sequentially, in that order.In another example, where the oil phase contains a non-polar solvent, apreservative, a co-surfactant, a surfactant and a non-polar compound,the non-polar solvent, the preservative and the co-surfactant are addedand mixed until the co-surfactant dissolves; the surfactant is addeduntil dissolved; and the non-polar active ingredient is added, in thatorder.

In one example, the methods for producing the concentrates are performedusing bench-top process, as provided herein. In another example,particularly when large batches of the concentrates, the methods areperformed with a scaled-up process, as described herein below, such asthe exemplary scaled-up process illustrated in FIG. 1.

In one example, where the polar solvent is water, the water is firstpurified by passage through purifiers. In one example, the waterpurification is carried out by passage through purifiers, sequentially,in the following order: a carbon filter, ion exchange equipment, reverseosmosis equipment, a 100 micron end-point filter, and a 50 micronpoint-of-use filter. In this example, after purification, the water isadded, with the other water phase ingredients, to a water phase tank.The water phase ingredient(s) then are mixed using a standard mixerattached to the tank, for example, mounted on the top of the tank. Aheating apparatus (typically the water jacket on the water phase tank)is used to heat the water phase ingredients during water phasegeneration, typically to low heat (e.g., 60° C.). To generate the oilphase, the oil phase ingredient(s) are weighed/measured and added to anoil phase tank. The oil phase ingredients are mixed using a standardmixer attached to the oil phase tank, for example, mounted on the tank.A heating apparatus (typically the water jacket on the oil phase tank)is used to heat the oil phase ingredients during water phase generation,typically to low heat (e.g., 60° C.). Once the oil and water phasesreach 60° C., and after oil phase components have dissolved, the oil andwater phases are combined by transferring the oil phase to the waterphase vessel, via transfer means. For this process, a homogenizermounted on the water phase tank, is turned on, for example, at 850-1200rpm. The ball valves then are opened and the transfer pump turned on,thereby effecting transfer of the oil phase liquid to the water phasetank via the transfer hose(s). As the phases are combined, the mixtureis homogenized by continued mixing with the homogenizer. The homogenizercan be adjusted, for example, by adjusting the baffle plate on thehomogenizer to achieve and maintain an emulsion, for example, by movingthe baffle plate further into the forming emulsion and/or further out ofthe forming emulsion. During the emulsifying step, the forming emulsionis cooled, typically rapidly cooled, by repeatedly passing the formingemulsion through a recirculating cooler, which is attached to the waterphase tank. The emulsion is transferred, via transfer means to aholding/packaging tank, where additional ingredients can be added and/orthe mixture can be evaluated. The additional ingredients are mixed intothe concentrate using a standard mixer. An end-product filter is used tofilter the concentrate before use.

Any of the provided methods for producing the concentrates can be usedto make any of the provided concentrates, as described herein.

Also provided are methods for producing the provided liquid dilutioncompositions containing the concentrates, such as beverages containingthe concentrates. These methods include methods for providing oil-basedadditives, for example, in a food or beverage. These methods includeadding any of the provided concentrates, e.g., liquid nanoemulsionconcentrates, to an aqueous medium, such as a beverage. Typically, theconcentrate is added to the medium, e.g., beverage, such that the mediumcontains an effective amount of the additive (e.g., the non-polar activeingredient).

The effective amount of the additive, such as the non-polar activeingredient is the quantity and/or concentration of the additivenecessary for preventing, curing, ameliorating, arresting or partiallyarresting a symptom of a disease or disorder, or the quantity and/orconcentration desired by an individual for intake, such as daily intake,and/or nutritional supplementation, for example, an amount sufficient toenhance the nutritional, pharmaceutical, nutraceutical, health or energyproperty of a food, beverage, or other consumable. In some examples, theconcentrate is added to the aqueous medium such that the resultingliquid dilution composition contains an effective amount of a particularnon-polar compound, for example, a particular amount per volume orweight of the composition, such as, for example, at least 25 mg or about25 mg, at least 35 mg or about 35 mg, at least 50 mg or about 50 mg, atleast 100 mg or about 100 mg, at least 250 mg or about 250 mg, or atleast 500 mg or about 500 mg of the non-polar active ingredient per 8fluid ounces of the liquid dilution composition.

In one example, an effective amount is a concentration or amount of theliquid nanoemulsion where at least 25 mg or about 25 mg, typically atleast 35 mg, for example, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220,230, 240, 250, 260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450,475, 500, 550, 600, 700, 800, 900, 1000, 1500, 2000 mg, or more, of thenon-polar active ingredient, is contained in at least 8 fluid ounces ofthe aqueous medium.

U.S. Provisional Application Ser. No. 61/070,381, filed Mar. 20, 2008,entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS;” and U.S.Provisional Application Ser. No. 61/132,424, filed Jun. 16, 2008,entitled “COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS,” each to PhilipBromley, for example, provide compositions containing non-polarcompounds and surfactants such as PEG-derivatives of Vitamin E, such asTocopherol Polyethylene glycol succinate (TPGS). Any of the compositionsdisclosed in the above-noted applications can be adapted to make theprovided compositions, for example, by replacing all or some of thePEG-derivatives of Vitamin E in the compositions with anothersurfactant(s), such as sugar esters, e.g., sucrose fatty acid esters,and other surfactants having similar HLB values, to make the providedcompositions.

The nanoemulsion concentrates are used to provide, by dilution,non-polar active ingredients to a beverage, such that a typical serving(i.e. between 4-10 ounces, for example, between 6-10 ounces,particularly about or 8 ounces) delivers a single dosage of suchingredient. The active ingredients of interest include, but are notlimited to, Omega-6 (CLA), Omega-3 (DHA/EPA), Vitamin D3, CoQ10 andPhytosterols. Typical single dosages of each can vary, but include forexample: Omega-6 (CLA) 1.5-3.0 grams per serving per day in, forexample, an 8 ounce beverage; Omega-3 DHA/EPA (DHA) 30-250 mg, such as32 or 50 mg, 100, 150, 200, 220 mg, Vitamin D3 200-1200 IU, 300-1000 IU,400-800 IU, such as 400, 500, 600, 700, 800, 900, 1000, 1200, 1500, 2000IU; CoQ10, 10-250 mg, 50-100 mg, 50-150 mg, 100-200 mg, such as 10, 50,100, 150, 200, 50-100 mg, 100-250 mg; and Phytosterols 100-1000 mg, such100, 200, 300, 400, 500, 600, 700, 800, 900 up to 1,000 mg per.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 sets forth an exemplary scaled-up process 100 of the providedmethods for making the liquid nanoemulsion concentrates. This process isexemplary and variations can be used. In this example, water 101 is usedas the polar solvent and the water first is purified by passage throughthe following purifiers, sequentially, in the following order: a carbonfilter 105, ion exchange equipment 106, reverse osmosis equipment 107, a100 micron end-point filter 108, and a 50 micron point-of-use filter109. After purification, the water is added, with the other water phaseingredients, to a water phase tank 103. In other examples, the polarsolvent is another polar solvent, such as glycerin or propylene glycol.In the illustrated example, the water phase ingredient(s) then are mixedusing a standard mixer 111 attached to the tank, for example, mounted onthe top of the tank. A heating apparatus (typically the water jacket onthe water phase tank) is used to heat the water phase ingredients duringwater phase generation, typically to low heat (e.g., 60° C.). Togenerate the oil phase, the oil phase ingredient(s) are weighed/measuredand added to an oil phase tank 102. The oil phase ingredients are mixedusing a standard mixer 111 attached to the oil phase tank, for example,mounted on the tank. A heating apparatus (typically the water jacket onthe oil phase tank) is used to heat the oil phase ingredients duringwater phase generation, typically to low heat (e.g., 60° C.). Once theoil and water phases reach 60° C., and after oil phase components havedissolved, the oil and water phases are combined by transferring the oilphase to the water phase vessel, via transfer means 112. For thisprocess, a homogenizer 110 mounted on the water phase tank, is turnedon, for example, at 850-1200 rpm. The ball valves then are opened andthe transfer pump turned on, thereby effecting transfer of the oil phaseliquid to the water phase tank via the transfer hose(s). As the phasesare combined, the mixture is homogenized by continued mixing with thehomogenizer 110. The homogenizer can be adjusted, for example, byadjusting the baffle plate on the homogenizer to achieve and maintain anemulsion, for example, by moving the baffle plate further into theforming emulsion and/or further out of the forming emulsion. During theemulsifying step, the forming emulsion is cooled, typically rapidlycooled, by repeatedly passing the forming emulsion through arecirculating cooler 115 (e.g., Model No. OC-1000 RO, sold by Turmoil,West Swanzey, N.H.), which is attached to the water phase tank. Theemulsion is transferred, via transfer means 112 to a holding/packagingtank 104, where additional ingredients can be added and/or the mixturecan be evaluated. The additional ingredients are mixed into theconcentrate using a standard mixer 111. An end-product filter 113 isused to filter the concentrate before use.

DETAILED DESCRIPTION

Outline

A. DEFINITIONS 28 B. COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS 63 1.Liquid nanoemulsion concentrates containing the non-polar compounds 65a. Formulating the liquid concentrates 67 i. Common ingredients andtypical concentration ranges 69 ii. Evaluation of the initialconcentrate 72 (1) Clarity 73 (2) Empirical evaluation 75 (3) Particlesize 75 (4) Turbidity measurement 76 iii. Selecting a formulation andmodifying formulations 78 b. Non-Polar Compounds 79 i. PolyunsaturatedFatty Acid (PUFA)-containing active ingredients 82 (1) Omega-3 fattyacid compounds 84 (a) DHA/EPA 84 (i) Fish Oils 85 (ii) Algae oil 87 (b)Flax Seed Oil - omega 3 (ALA) 88 (2) Omega-6 compounds 89 (a) Borage oil(Gamma-Linolenic Acid (GLA)) 89 (3) Saw Palmetto extract 90 (4)Conjugated Linoleic Acid (CLA) 90 ii. Coenzyme Q Active Ingredients 91(1) Coenzyme Q10 91 iii. Phytosterol-Containing Active Ingredients 92 c.Surfactants 93 (1) Sucrose Fatty Acid Ester Surfactants 95 (2)Production of Sucrose Esters 102 ii. Concentration of the surfactant 111iii. HLB 113 d. Co-surfactants (emulsifiers) 114 i. Phospholipids 114 e.Polar solvents 115 f. Preservatives and Sterilizers 118 g. Emulsionstabilizers (co-emulsifier) 119 h. Non-polar solvents 120 i. Flavors 121j. pH adjusters 121 2. Powder forms of the compositions 122 3. Liquiddilution compositions containing the diluted concentrates 126 a. Clarity128 i. Clarity determined by empirical evaluation 128 ii. Claritydetermined by particle size or number of particles 129 iii. Turbidity129 b. Stability 130 c. Desirable characteristics for human consumption132 d. Safety 132 e. Oral bioavailability 132 C. METHODS FOR MAKINGLIQUID NANOEMULSION CONCENTRATES 133 CONTAINING NON-POLAR COMPOUNDS 1.Equipment for making the concentrates 133 2. Scales 133 a. Purifiers,including filters 134 b. Vessels for mixing the ingredients 135 c.Mixers 136 d. Heating apparatuses 138 e. Cooling apparatuses 138 f.Transfer means 139 g. Evaluation equipment 140 3. General methods formaking the liquid nanoemulsion concentrates 140 a. Generating the waterphase 142 i. Water phase ingredients 143 b. Generating the oil phase 144i. Oil phase ingredients 145 c. Combining and emulsifying the oil phaseand the water phase 146 i. Combining the oil and water phases 146 ii.Emulsifying the oil and water phases 147 iii. Cooling 147 d. Additionalsteps 148 i. Additional ingredients 149 ii. Evaluation of theconcentrate 149 iii. Filtering the concentrate 149 4. Bench-top process149 5. Scaled-up manufacturing processes 151 a. Water purification 152b. Generation of the water phase and oil phase: 152 c. Combining andemulsifying the phases 153 d. Cooling 154 e. Additional steps 154 D.METHODS FOR MAKING THE LIQUID DILUTION COMPOSITIONS 155 CONTAINING THEDILUTED CONCENTRATES 1. Dilutions 156 2. Analyzing the aqueous liquiddilution compositions containing the liquid 157 concentrates a.Clarity/turbidity 157 i. Empirical evaluation 158 ii. Particle size 158iii. Turbidity measurement 159 E. EXAMPLES 160A. Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which the invention(s) belong. All patents, patent applications,published applications and publications, GENBANK sequences, websites andother published materials referred to throughout the entire disclosureherein, unless noted otherwise, are incorporated by reference in theirentirety. In the event that there is a plurality of definitions forterms herein, those in this section prevail. Where reference is made toa URL or other such identifier or address, it is understood that suchidentifiers can change and particular information on the internet cancome and go, but equivalent information is known and can be readilyaccessed, such as by searching the internet and/or appropriatedatabases. Reference thereto evidences the availability and publicdissemination of such information.

As used herein, colloid refers to a mixture containing two phases, adispersed phase and a continuous phase, the dispersed phase containingparticles (droplets) distributed throughout the continuous phase.Colloidal mixtures include aerosols, foams and dispersions, for example,emulsions, for example, nanoemulsions. A liquid colloid, for example, ananoemulsion, can have a similar appearance, for example, clarity, to asolution, in which there is no dispersed phase.

As used herein, emulsion refers to a colloidal dispersion of twoimmiscible liquids, for example, an oil and water (or other aqueousliquid, e.g., a polar solvent), one of which is part of a continuousphase and the other of which is part of a dispersed phase. The providedcompositions include emulsions, typically oil-in-water nanoemulsions(which include any oil soluble phase dispersed in any aqueous phase,also called the water phase), in which the oil phase is the dispersedphase and the water phase is the continuous phase. Emulsions typicallyare stabilized by one or more surfactants and/or co-surfactants and/oremulsion stabilizers. Surfactants form an interfacial film between theoil and water phase of the emulsion, providing stability. Typically, thenanoemulsions of the provided compositions contain micelles, containingone or more surfactant surrounding a non-polar active ingredient, whichare dispersed in the water phase. Exemplary of the provided emulsionsare the provided liquid nanoemulsion concentrates and liquid dilutioncompositions made by diluting the concentrates, typically in an aqueousmedium.

As used herein, a nanoemulsion is an emulsion in which the disperseddroplets, for example, the micelles, have a diameter (particle size)less than 1000 nm or less than about 1000 nm, typically, less than 500nm or less than about 500 nm, typically less than 300 nm or about 300nm, for example, less than 250 nm or about 250 nm, for example, lessthan 200 nm or less than about 200 nm, for example, less than or lessthan about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130,140, 150, 160, 170, 180, 190, or 200 nm. Exemplary of nanoemulsions arethe provided liquid nanoemulsion concentrates and the liquid dilutioncompositions, for example, the aqueous liquid dilution compositionscontaining the diluted concentrates.

As used herein, “surfactant” and “surface active agent” refersynonymously to synthetic and naturally occurring amphiphilic moleculesthat have hydrophobic portion(s) and hydrophilic portion(s). Due totheir amphiphilic (amphipathic) nature, surfactants and co-surfactantstypically can reduce the surface tension between two immiscible liquids,for example, the oil and water phases in an emulsion, stabilizing theemulsion. Different surfactants can be characterized based on theirrelative hydrophobicity and/or hydrophilicity. For example, relativelylipophilic surfactants are more soluble in fats, oils and waxes,typically having HLB values less than 10 or about 10, while relativelyhydrophilic surfactants are more soluble in aqueous compositions, forexample, water, and typically have HLB values greater than 10 or about10. Relatively amphiphilic surfactants are soluble in oil and waterbased liquids and typically have HLB values close to 10 or about 10.

Surfactants include, for example, soaps, detergents, lipids,emulsifiers, dispersing agents and wetting agents, molecules thatemulsify liquids, for example, by forming an emulsion in an aqueousmedium or aqueous liquid dilution composition, for example, forming acolloidal dispersion of two immiscible liquids in the form of droplets,for example, an emulsion such as a microemulsion; and compounds thatform various macromolecular structures, for example, aggregates, inliquids, for example, micelles, lipid bilayer structures, includingliposomes, and inverse micelles.

Typically, the surfactants used in the provided compositions have an HLBvalue between 14 or about 14 and 20 or about 20, for example, at orabout 14, 15, 16, 17, 18, 19 or 20, and typically between at of about 15and at or about 18. Exemplary of the surfactants include, but are notlimited to, non-ionic surfactants, such as sugar ester surfactants, suchas sucrose fatty acid ester (SFAE) surfactants, which typically includesucrose fatty acid monoesters, such as sucrose monolaurate, sucrosemonopalmitate, sucrose monostearate, and sucrose monooleate, and allmixtures of a SFAE(s) and a PEG-derivative of vitamin E, such as TPGS oran analog thereof. Typically, the surfactant is a natural surfactant,for example, a surfactant that is G.R.A.S. (generally recognized assafe) by the FDA and/or Kosher certified.

As used herein, analog refers to a chemical compound that isstructurally similar to another compound (referred to as a parentcompound), but differs slightly in composition, for example, by thevariation, addition or removal of an atom, one or more units (e.g.,methylene unit(s), —(CH₂)_(n)—) or one or more functional groups. Theanalog can have different chemical or physical properties compared withthe original compound and/or can have improved biological and/orchemical activity. Alternatively, the analog can have similar oridentical chemical or physical properties compared with the originalcompound and/or can have similar or identical biological and/or chemicalactivity. For example, the analog can be more hydrophilic or it can havealtered reactivity as compared to the parent compound. The analog canmimic the chemical and/or biologically activity of the parent compound(i.e., it can have similar or identical activity), or, in some cases,can have increased or decreased activity. The analog can be a naturallyor non-naturally occurring (e.g., synthetic) variant of the originalcompound. Other types of analogs include isomers (e.g., enantiomers,diastereomers) and other types of chiral variants of a compound, as wellas structural isomers. The analog can be a branched or cyclic variant ofa linear compound. For example, a linear compound can have an analogthat is branched or otherwise substituted to impart certain desirableproperties (e.g., improve hydrophobicity or bioavailability). Exemplaryof the analogs used in the provided compositions and methods are sucrosefatty acid ester analogs, which can be used as surfactants in place ofthe sucrose fatty acid ester parent compound surfactants in the providedcompositions.

As used herein, homolog refers to an analog that differs from the parentcompound only by the presence or absence of a simple unit, such as amethylene unit, or some multiple of such units, e.g., —(CH₂)_(n)—.Typically, a homolog has similar chemical and physical properties as theparent compound. Exemplary of the homologs used in the providedcompositions and methods are sucrose fatty acid ester homologs.

As used herein, a PEG derivative of Vitamin E is a compound containingone or more Vitamin E moiety (e.g., a tocopherol or tocotrienol) joined,for example by an ester, ether amide or thioester bond, with one or morepolyethylene glycol (PEG) moieties, via a linker, for example adicarboxylic or tricarboxylic acid. Exemplary of PEG derivatives ofVitamin E are tocopherol polyethylene glycol succinate (TPGS), TPGSanalogs, TPGS homologs and TPGS derivatives.

As used herein, a tocopherol polyethylene glycol diester (TPGD) is aPEG-derivative of tocopherol where the linker is a dicarboxylic acid (acarboxylic acid having two carboxy groups, e.g., succinic acid), such assuccinic acid. Exemplary of dicarboxylic acids that can be used aslinkers in these tocopherol and tocotrienol PEG diester surfactants aresuccinic acid, sebacic acid, dodecanedioic acid, suberic acid, orazelaic acid, citraconic acid, methylcitraconic acid, itaconic acid,maleic acid, glutaric acid, glutaconic acid, fumaric acids and phthalicacids. Exemplary of TPGDs are tocopherol succinate polyethylene glycol(TPGS), tocopherol sebacate polyethylene glycol, tocopheroldodecanodioate polyethylene glycol, tocopherol suberate polyethyleneglycol, tocopherol azelaate polyethylene glycol, tocopherol citraconatepolyethylene glycol, tocopherol methylcitraconate polyethylene glycol,tocopherol itaconate polyethylene glycol, tocopherol maleatepolyethylene glycol, tocopherol glutarate polyethylene glycol,tocopherol glutaconate polyethylene glycol, and tocopherol phthalatepolyethylene glycol, among others.

As used herein, “tocopherol polyethylene glycol succinate” “TPGS,”“tocopheryl polyethylene glycol succinate surfactant” and “TPGSsurfactant” refer to tocopherol polyethylene glycol (PEG) diesters, thatare formed by joining, via esterification, tocopherol succinate, whichitself is an ester made by esterification of tocopherol and succinicacid. The term tocopherol refers to any naturally occurring or syntheticform of vitamin E, and can refer to a single compound or a mixture.Examples of tocopherols include, for example, α-tocopherol,D-α-tocopherol, β-tocopherol, γ-tocopherol, and δ-tocopherol. The PEGmoiety of the TPGS surfactant can be any PEG moiety, for example, PEGmoieties between 200 kDa or about 200 kDa and 20,000 kDa or about 20,000kDa, typically between 200 kDa or about 200 kDa and 6000 kDa or about6000 kDa, for example, between 600 kDa or about 600 kDa and 6000 kDa orabout 6000 kDa, typically between 200 kDa or about 200 kDa and 2000 kDaor about 2000 kDa, between 600 kDa or about 600 kDa and 1500 kDa orabout 1500 kDa, or between 600 kDa or about 600 kDa and 1000 kDa orabout 1000 kDa, for example, 200 kDa or about 200 kDa, 300 kDa or about300 kDa, 400 kDa or about 400 kDa, 500 kDa or about 500 kDa, 600 kDa orabout 600 kDa, 800 kDa or about 800 kDa, and 1000 kDa or about 1000 kDa;and PEG moieties that are modified, for example, methylated PEG (m-PEG))and/or PEG moieties including other PEG analogs, e.g., PEG-NHS,PEG-aldehyde, PEG-SH, PEG-NH₂, PEG-CO₂H, and branched PEGs.

Exemplary of the TPGS surfactants is TPGS-1000, which has a PEG moietyof 1000 kDa. The TPGS can be any natural, water-soluble, tocopherolpolyethylene glycol succinate, for example, the food grade TPGS soldunder the name Eastman Vitamin E TPGS®, food grade, by Eastman ChemicalCompany, Kingsport, Tenn. This TPGS is water-soluble form ofnatural-source vitamin E, which is prepared by esterifying the carboxylgroup of crystalline d-alpha-tocopheryl acid succinate with polyethyleneglycol 1000 (PEG 1000), and contains between 260 and 300 mg/g totaltocopherol. A similar compound can be made by esterifying the carboxylgroup of the d,1 form of synthetic Vitamin E with PEG 1000. It forms aclear liquid when dissolved 20% in water. This tocopheryl polyethyleneglycol is a water-soluble preparation of a fat-soluble vitamin (vitaminE), for example, as disclosed in U.S. Pat. Nos. 3,102,078, 2,680,749 andU.S. Published Application Nos. 2007/0184117 and 2007/0141203. Alsoexemplary of the TPGS surfactant that can be used in the providedcompositions is the Water Soluble Natural Vitamin E (TPGS), sold byZMC-USA, The Woodlands, Tex. Any known source of TPGS can be used.Typically, the TPGS surfactant is GRAS and Kosher certified. TPGStypically has an HLB value of between 16 or about 16 and 18 or about 18.

As used herein, analog refers to a chemical compound that isstructurally similar to another compound (referred to as a parentcompound), but differs slightly in composition, for example, by thevariation, addition or removal of an atom, one or more units (e.g.,methylene unit(s), —(CH₂)_(n)—) or one or more functional groups. Theanalog can have different chemical or physical properties compared withthe original compound and/or can have improved biological and/orchemical activity. Alternatively, the analog can have similar oridentical chemical or physical properties compared with the originalcompound and/or can have similar or identical biological and/or chemicalactivity. For example, the analog can be more hydrophilic or it can havealtered reactivity as compared to the parent compound. The analog canmimic the chemical and/or biologically activity of the parent compound(i.e., it can have similar or identical activity), or, in some cases,can have increased or decreased activity. The analog can be a naturallyor non-naturally occurring (e.g., synthetic) variant of the originalcompound. Other types of analogs include isomers (e.g., enantiomers,diastereomers) and other types of chiral variants of a compound, as wellas structural isomers. The analog can be a branched or cyclic variant ofa linear compound. For example, a linear compound can have an analogthat is branched or otherwise substituted to impart certain desirableproperties (e.g., improve hydrophobicity or bioavailability). Exemplaryof the analogs used in the provided compositions and methods are TPGSanalogs, which can be used as surfactants in place of the TPGS in theprovided compositions.

As used herein, homolog refers to an analog that differs from the parentcompound only by the presence or absence of a simple unit, such as amethylene unit, or some multiple of such units, e.g., —(CH₂)_(n)—.Typically, a homolog has similar chemical and physical properties as theparent compound. Exemplary of the homologs used in the providedcompositions and methods are TPGS homologs.

As used herein, “tocopherol polyethylene glycol succinate analog”, “TPGSanalog”, and “TPGS analog surfactant” refer to compounds, other thanTPGS, that are similar to a parent TPGS compound, but differ slightly incomposition, for example, by the variation, addition or removal of anatom, one or more units (e.g., methylene unit(s)—(CH₂)_(n)) or one ormore functional groups. TPGS analogs include Vitamin E derivedsurfactants, including PEG derivatives of Vitamin E, including vitamin EPEG diesters, such as, but not limited to, tocopherol polyethyleneglycol sebacate (PTS), tocopherol polyethylene glycol dodecanodioate(PTD), tocopherol polyethylene glycol suberate (PTSr), tocopherolpolyethylene glycol azelaate (PTAz), and polyoxyethanyl tocotrienylsebacate (PTrienS) as well as other PEG derivatives of Vitamin E. In oneexample, the surfactant in the provided compositions is a TPGS analog.

Exemplary of TPGS analogs are compounds, other than TPGS compounds,having the formula shown in Scheme I:

where R¹, R², R³ and R⁴ each independently is hydrogen (H) or methyl(CH₂); each dashed line is, independently, a single or double bond; n isan integer from 1 to 5000; m and q each independently are 0 or 1; and pis an integer from 1 to 20.

For example, TPGS analogs include, but are not limited to, compoundshaving the formula in Scheme I, where, when the bonds represented by thedashed lines marked by “A” and “B” are single bonds, and each of m and qequals 0, p is any integer from 2-20. TPGS analogs also includecompounds where the dashed line at B or the dashed line at A, or boththe dashed lines, represents at least one double bond. For example, TPGSanalogs include a compound as in Scheme I, where when the dashed line inA represents only single bonds, the dashed line in “B” represents one ormore double bond, e.g., tocotrienol PEG diesters. TPGS analogs alsoinclude compounds as in Scheme I, where when the dashed line marked “B”represents only single bonds, the dashed line marked “A” represents oneor more double bonds; or when the dashed line labeled “A” does notrepresent double bonds, and each of m and q is zero, p is greaterthan 1. For example, TPGS analogs include compounds where one or more ofthe dashed lines represents a double bond, for example, PEG derivativesof tocotrienol esters (e.g., PTrienS).

Also exemplary of TPGS analogs are compounds, other than TPGS compounds,having the formula shown in Scheme II:

where R¹, R², R³ and R⁴ each independently is hydrogen (H) or methyl(CH₂); the bond represented by the dashed line is either a single ordouble bond; and m is a integer from 1 to 20, and n is an integer from 1to 5000.

Also exemplary of TPGS analogs include compounds other than TPGS, havingPEG moieties that vary in chain length, according to the formula shownin Scheme III:

where R¹, R², R³ and R⁴ each independently is hydrogen (H) or methyl(CH₂), and n is an integer from 1 to 5000.

As used herein, TPGS-1000 analogs are compounds other than TPGS-1000that are similar to a parent TPGS-1000 compound, but differ slightly incomposition, for example, by the variation, addition or removal of anatom, one or more units (e.g., methylene unit(s)—(CH₂)_(n)) or one ormore functional groups. In one example, the surfactant in thecompositions provided herein is a TPGS-1000 analog. Suitable TPGS-1000analogs include, but are not limited to, other TPGS compounds, havingPEG moiety(ies) that vary in chain length and molecular weight comparedto TPGS-1000, including, for example, TPGS compounds having PEG moietiesbetween 200 or about 200 kDa and 20,000 kDa or about 20,000 kDa,typically between 200 kDa or about 200 kDa and 6000 kDa or about 6000kDa, for example, between 600 kDa or about 600 kDa and 6000 kDa or about6000 kDa, typically between 200 kDa or about 200 kDa and 2,000 kDa orabout 2,000 kDa, between 600 kDa or about 600 kDa and 1500 kDa or about1500 kDa, such as, but not limited to, 200, 300, 400, 500, 600, 800, and1000 kDa. Also exemplary of TPGS-1000 analogs are TPGS compounds havingPEG moieties that are modified, for example, methylated PEG (m-PEG)and/or PEG moieties including other PEG analogs, e.g., PEG-NHS,PEG-aldehyde, PEG-SH, PEG-NH₂, PEG-CO₂H, and branched PEGs. Alsoexemplary of TPGS-1000 analogs are any TPGS analogs, e.g., Vitamin Ederived surfactants, including PEG derivatives of Vitamin E, includingvitamin E PEG diesters, such as, but not limited to, tocopherolpolyethylene glycol sebacate (PTS), tocopherol polyethylene glycoldodecanodioate (PTD), tocopherol polyethylene glycol suberate (PTSr),tocopherol polyethylene glycol azelaate (PTAz) and polyoxyethanyltocotrienyl sebacate (PTrienS) as well as other PEG derivatives ofVitamin E.

As used herein, TPGS homologs are analogs of TPGS that differ from aTPGS parent compound only by the presence or absence of a simple unit,such as a methylene unit, or some multiple of such units, e.g.,—(CH₂)_(n)—. In one aspect, TPGS homologs are used as surfactants in theprovided compositions. Typically, suitable TPGS homologs have similarsurfactant properties compared to the parent compound (TPGS), forexample, similar HLB values, for example, HLB values between 14 or about14 and 20 or about 20. Exemplary of TPGS homologs are tocopherolpolyethylene glycol sebacate (PTS), tocopherol polyethylene glycoldodecanodioate (PTD), tocopherol polyethylene glycol suberate (PTSr),tocopherol polyethylene glycol azelaate (PTAz). Exemplary of TPGShomologs are compounds having the formula in Scheme I (above), whereneither the A or B dashed line represents a double bond and where, wheneach of m and q is 0, p is greater than 1.

As used herein, TPGS-1000 homologs are analogs of TPGS-1000 that differfrom a TPGS-1000 parent compound only by the presence or absence of asimple unit, such as a methylene unit, or some multiple of such units,e.g., —(CH₂)_(n)—. Suitable TPGS-1000 homologs have similar surfactantproperties compared to the parent compound (TPGS-1000), for example,similar HLB values, for example, HLB values between 14 or about 14 and20 or about 20. Suitable TPGS-1000 homologs include TPGS-1000 homologswith slight variations in the length of the PEG chain moiety, andme-TPGS-1000, which is a TPGS-1000 having a methyl cap on the PEGmoiety.

As used herein, HLB refers to a value that is used to index and describea surfactant according to its relative hydrophobicity/hydrophilicity,relative to other surfactants. A surfactant's HLB value is an indicationof the molecular balance of the hydrophobic and lipophilic portions ofthe surfactant, which is an amphipathic molecule. Each surfactant andmixture of surfactants (and/or co-surfactants) has an HLB value that isa numerical representation of the relative weight percent of hydrophobicand hydrophilic portions of the surfactant molecule(s). HLB values arederived from a semi-empirical formula. The relative weight percentagesof the hydrophobic and hydrophilic groups are indicative of surfactantproperties, including the molecular structure, for example, the types ofaggregates the surfactant will form and the solubility of thesurfactant. See, for example, Griffin, W. C. J. Soc. Cos. Chem. 1:311(1949).

Surfactant HLB values range from 1-45, while the range for non-ionicsurfactants typically is from 1-20. The more lipophilic a surfactant is,the lower its HLB value. Conversely, the more hydrophilic a surfactantis, the higher its HLB value. Lipophilic surfactants have greatersolubility in oil and lipophilic substances, while hydrophilicsurfactants dissolve more easily in aqueous media. In general,surfactants with HLB values greater than 10 or greater than about 10 arecalled “hydrophilic surfactants,” while surfactants having HLB valuesless than 10 or less than about 10 are referred to as “hydrophobicsurfactants.” HLB values have been determined and are available for aplurality of surfactants (e.g., see U.S. Pat. No. 6,267,985). It shouldbe appreciated that HLB values for a given surfactant or co-surfactantcan vary, depending upon the empirical method used to determine thevalue. Thus, HLB values of surfactants and co-surfactants provide arough guide for formulating compositions based on relativehydrophobicity/hydrophilicity. For example, a surfactant typically isselected from among surfactants having HLB values within a particularrange of the surfactant or co-surfactant that can be used to guideformulations. Table 1A lists HLB values of exemplary surfactants andco-surfactants. Table 1B (see section B(1)(c) for exemplary sucrosefatty acid ester surfactants and their HLB values).

TABLE 1A HLB Values of Exemplary Surfactants and Co-SurfactantsSurfactant/ Surfactant/ co-surfactant HLB co-surfactant HLB PEG-2Hydrogenated 1.7 PEG-10 oleyl ether 12.4 Castor Oil Sorbitan Trioleate1.8 PEG-8 isooctylphenyl ether 12.4 Sorbitan Tristearate 2.1 PEG-10stearyl ether 12.4 Glyceryl Stearate 3.5 PEG-35 Castor Oil 12.5 SorbitanSesquioleate 3.7 PEG-10 cetyl ether 12.9 Labrafil 4 Nonoxynol-9 12.9Sorbitan Oleate 4.3 PEG-40 Castor Oil 13 Sorbitan monostearate 4.7PEG-10 isooctylphenyl ether 13.5 PEG-2 oleyl ether 4.9 PEG-40Hydrogenated Castor Oil 14 PEG-2 stearyl ether 4.9 Labrasol 14 PEG-7Hydrogenated 5 Nonoxynol-15 14.2 Castor Oil PEG-2 cetyl ether 5.3 PEG-12tridecyl ether 14.5 PEG-4 Sorbitan 5.5 PEG-18 tridecyl ether 14.5Stearate PEG-2 Sorbitan 6 Polysorbate 60 14.9 Isostearate SorbitanPalmitate 6.7 Polysorbate 80 15 Triton SP-135 8 PEG-20 Glyceryl Stearate15 Sorbitan monolaurate 8.6 PEG-20 Stearate 15 PEG-40 Sorbitan 9.5PEG-20 stearyl ether 15.3 Peroleate PEG-4 lauryl ether 9.7 PEG-20 oleylether 15.3 Polysorbate 81 10 Polysorbate 40 15.6 PEG-40 Sorbitan 10PEG20 cetyl ether 15.7 Hexaoleate PEG-40 Sorbitan 10 PEG(20) hexadecylether 15.7 Perisostearate PEG-10 Olive 10 PEG-60 Hydrogenated Castor Oil16 Glycerides PEG sorbitol 10.2 PEG-30 Stearate 16.5 hexaoleatePolysorbate 65 10.5 Polysorbate 20 16.7 PEG-25 Hydrogenated 10.8 PEG-75Lanolin 16.7 Castor Oil Polysorbate 85 11 PEG23 lauryl ether 16.9 PEG-7Glyceryl 11 PEG-40 Stearate 17.3 Cocoate PEG-8 Stearate 11.1 PEG-50Stearate 17.7 PEG sorbitan 11.4 PEG40 isooctylphenyl ether 17.9tetraoleate PEG-15 Glyceryl 12 PEG-100 Stearate 18.8 Isostearate PEG-35Almond 12 Pluronic F68 29 Glycerides Tocopherol 16-18Phosphatidylcholine 7.6 polyethylene glycol succinate (TPGS)

The surfactants and HLB values set forth in Table 1A and Table 1B(below, see section B(1)(c)) are exemplary. Any known surfactant orco-surfactant can be used with the provided compositions (see, e.g.,U.S. Pat. No. 6,267,985). The surfactant(s) contained in the providedcompositions typically have an HLB value between 14 or about 14 and 20or about 20, for example, 14 or about 14, 15 or about 15, 16 or about16, 17 or about 17, 18 or about 18, 19 or about 19, and 20 or about 20.

As used herein, micelle refers to aggregates formed by surfactants thattypically form when the surfactant is present in an aqueous composition,typically when the surfactant is used at a concentration above thecritical micelle concentration (CMC). In micelles, the hydrophilicportions of the surfactant molecules contact the aqueous or the waterphase, while the hydrophobic portions form the core of the micelle,which can encapsulate non-polar ingredient(s), for example, thenon-polar compounds in the provided compositions. Typically, thesurfactants in the provided compositions form micelles containing thenon-polar ingredient at their center in aqueous liquid dilutioncompositions. Typically, the micelles in the provided compositions havea particle size of about 1000 nm, typically, less than 500 nm or lessthan about 500 nm, typically less than 300 or about 300 nm, for example,less than 250 nm or about 250 nm, for example, less than 200 nm or lessthan about 200 nm, for example, less than or less than about 5, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180,190, or 200 nm.

As used herein, inverse micelles are surfactant aggregates thattypically form in lipophilic solution, with the hydrophilic portionsforming the core. When the cross sectional area of the hydrophobicregion of the surfactant molecule is greater than that of thehydrophilic part of the molecule, the formation of micelles, which canbe hexagonal phase structures, is favored.

As used herein, liposomes are surfactant aggregates composed of lipidbilayers, typically having an aqueous core. Liposomes typically areformed by lipid surfactants, typically, phospholipids, which areamphipathic, phosphate-containing lipids, for example, moleculescontaining one phosphate, a glycerol and one or more fatty acids, andsimilar surfactants. Alternatively, phospholipid surfactants can be usedas co-surfactants, which can be incorporated into aggregates of othersurfactant(s), for example, micelles. Lipid bilayers are two dimensionalsheets in which all of the hydrophobic portions, e.g., acyl side chains,are shielded from interaction with aqueous liquid, except those at theends of the sheet. An energetically unfavorable interaction of the acylchains with water results in the folding of the bilayers to formliposomes, three-dimensional lipid bilayer vesicles. In one example, theliposome is formed as a single bilayer enclosing a single aqueous space(small unilamellar vesicles; SUVS). In another example, the liposome iscomposed of concentric bilayers with many aqueous spaces alternatingwith the bilayers (multilamellar vesicles; MLVS). Liposomes can be usedto encapsulate hydrophobic and hydrophilic active ingredients. Inliposomes, non-polar active ingredients typically are partitioned withinthe bilayers whereas hydrophilic active ingredients typically aretrapped within the aqueous compartments. In one example, liposomes canbe advantages as a carrier/encapsulation system because they are stableand can protect the active ingredients from degradation, e.g., by oxygenand digestive enzymes.

As used herein, “co-surfactant” is used to refer to a surfactant,typically a phospholipid, that is used, in the provided compositions, incombination with a surfactant (e.g., a primary surfactant), for example,to improve the emulsification of the provided compositions and/orcompounds, for example, to emulsify the ingredients. In one example, theprovided compositions contain at least one surfactant and at least oneco-surfactant. Typically, the co-surfactant is a lipid, for example, aphospholipid, for example, phosphatidylcholine. In one example, theco-surfactant has an HLB value of between 7 or about 7 and 8 or about 8.Typically, the co-surfactant represents a lower percent, by weight(w/w), of the provided compositions, compared to the surfactant. Thus,the provided compositions typically have a lower concentration of theco-surfactant(s) than of the surfactant.

As used herein, a phospholipid is an amphipathic, phosphate-containinglipid, for example, a molecule containing one phosphate, a glycerol andone or more fatty acids. In one example, one or more phospholipids isused as a co-surfactant in the provided compositions. Exemplary of thephospholipids used in the provided compositions are lecithin, includingphosphatidylcholine (PC), phosphatidylethanolamine (PE),distearoylphosphatidylcholine (DSPC), phosphatidylserine (PS),phosphatidylglycerol (PG), phosphatidic acid (PA), phosphatidylinositol(PI), sphingomyelin (SPM) or a combination thereof. Typically, thephospholipid is phosphatidylcholine (PC), which sometimes is referred toby the general name “lecithin.” Exemplary of the phospholipids that canbe used as co-surfactants in the provided compositions are thephospholipids sold by Lipoid, LLC, Newark, N.J., for example, PurifiedEgg Lecithins, Purified Soybean Lecithins, Hydrogenated Egg and SoybeanLecithins, Egg Phospholipids, Soybean Phospholipids, Hydrogenated Eggand Soybean Phospholipids. Synthetic Phospholipids, PEG-ylatedPhospholipids and phospholipid blends sold by Lipoid, LLC. Exemplary ofthe phosphatidylcholine that can be used as a co-surfactant in theprovided compositions is the phosphatidylcholine composition sold byLipoid, LLC, under the name Lipoid S100, which is derived from soyextract and contains greater than 95% or greater than about 95%phosphatidylcholine.

Typically, for micelle formation, surfactant(s) are used in which thecross sectional area of the hydrophilic portion of the surfactantmolecule is greater than that of the hydrophobic portion of themolecule. For example, sucrose fatty acid ester surfactants having anHLB within the range of between at or about 14 and at or about 20,typically between at or about 15 and at or about 18, are surfactants forstabilizing oil-in-water emulsions containing the non-polar activeingredients, for example, in nanometer-sized droplets suspended ordispersed in an aqueous phase or aqueous liquid, for example, aqueousmedium, as spherical micelles, containing the hydrophilic portions ofthe molecule(s) facing the aqueous phase and the hydrophobic portions atthe center of the spherical micelles, for example, surrounding thenon-polar active ingredient. Typically, the surfactants and/orco-surfactants in the provided compositions aggregate in thenanoemulsions and the aqueous liquids to form micelles, which containthe non-polar compound(s). The hydrophilic portion(s) of the surfactantmolecules are oriented toward the outside of the micelle, in contactwith the aqueous medium, while the hydrophobic portion(s) of thesurfactant molecules are oriented toward the center of the micelle, incontact with the non-polar compound(s), which is contained in the centerof the micelle. The micelles can contain more than one surfactant.

As used herein, “particle size” and “average particle size” refersynonymously to the average diameter of particles in a provided liquid,for example, the droplet diameter or micelle diameter in an emulsion.Typically, the provided nanoemulsion concentrates, and the liquids madefrom the concentrates, have a particle size of less than about 1000 nm,typically, less than 500 nm or less than about 500 nm, typically lessthan 300 nm or about 300 nm, for example, less than 250 nm or about 250nm, for example, less than 200 nm or less than about 200 nm, forexample, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80,90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm. In oneexample, the dilution compositions yielded by diluting the liquidnanoemulsion concentrates have a particle size between 10 nm or about 10nm and 1000 nm or about 1000 nm, for example, between 15 nm or about 15nm and 500 nm or about 500 nm, for example, between 15 nm or about 15 nmand 300 nm or about 300 nm, for example, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120,130, 140, 150, 160, 170, 180, 190, 200 nm or more. Typically, theprovided liquid nanoemulsion concentrates are formulated such that,dilution of the liquid nanoemulsion concentrates in an aqueous mediumyields a liquid dilution composition having an appropriate particlesize, for example, between 15 nm or about 15 nm and 500 nm or about 500nm. Information about particles in the liquid dilution compositions,alternatively, can be expressed in terms of particle density, forexample, ppm (parts per million) or percent solids, in the liquids.

As used herein, visible particles are particles, for example, in aliquid, for example, an emulsion, that are visible when viewing theliquid with the naked eye (e.g., without magnification). In one example,the visible particles are particles that are observed by the artisanformulating the compositions, for example, the concentrates or theaqueous liquid dilution compositions containing the dilutedconcentrates. In one example, the provided compositions contain novisible particles. In another example, the compositions contain fewvisible particles, for example, no more visible particles than anotherliquid, for example, a beverage. The presence of visible particles andthe number of visible particles is determined by empirical observation.

As used herein, visible crystals are crystals that are visible whenviewing a liquid with the naked eye (e.g., without magnification). Thepresence or absence of visible crystals typically is determined byempirical observation and can be observed by the artisan formulating thecompositions, for example, the concentrates or the aqueous liquiddilution compositions containing the diluted concentrates. In oneexample, the provided compositions contain no visible crystals. Inanother example, the compositions contain few visible crystals, forexample, no more visible crystals than are contained in another liquid,for example, a beverage.

As used herein, “turbidity” is a measure of the cloudiness or hazinessof a liquid, caused by particles in suspension in the liquid. Turbiditycan measured optically, for example, using a nephelometer, an instrumentwith a light and a detector. The nephelometer measures turbidity bydetecting scattered light resulting from exposure of the liquid to anincident light. The amount of scattered light correlates to the amountof particulate matter in the liquid. For example, a beam of light willpass through a sample with low turbidity with little disturbance. Othermethods for measuring turbidity are well known and can be used with theprovided methods and compositions. The units of a turbidity valuemeasured with a nephelometer are Nephelometric Turbidity Units (NTU). Inone example, the provided compositions, e.g., the aqueous liquiddilution compositions containing the diluted liquid nanoemulsionconcentrates, have low turbidity, for example, a turbidity value (NTU)of 30 or about 30; or an NTU value of less than 30 or about 30, forexample, less than 29 or about 29, less than 28 or about 28, less than27 or about 27, less than 26 or about 26, less than 25 or about 25, lessthan 24 or about 24, less than 23 or about 23, less than 22 or about 22,less than 21 or about 21, less than 20 or about 20, less than 19 orabout 19, less than 18 or about 18, less than 17 or about 17, less than16 or about 16, less than 15 or about 15, less than 14 or about 14, lessthan 13 or about 13, less than 12 or about 12, less than 11 or about 11,less than 10 or about 10, less than 9 or about 9, less than 8 or about8, less than 7 or about 7, less than 6 or about 6, less than 5 or about5, less than 4 or about 4, less than 3 or about 3, less than 2 or about2, less than 1 or about 1; or 29 or about 29, 28 or about 28, 27 orabout 27, 26 or about 26, 25 or about 25, 24 or about 24, 23 or about23, 22 or about 22, 21 or about 21, 20 or about 20, 19 or about 19, 18or about 18, 17 or about 17, 16 or about 16, 15 or about 15, 14 or about14, 13 or about 13, 12 or about 12, 11 or about 11, 10 or about 10, 9 orabout 9, 8 or about 8, 7 or about 7, 6 or about 6, 5 or about 5, 4 orabout 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0 or about 0. Inanother example, the turbidity value of the aqueous liquid dilutioncomposition is less than 1000 or less than about 1000, less than 500 orless than about 500, less than 300 or less than about 300, less than 250or less than about 250, 200 or less than about 200, for example, 200,175, 150, 100, 50, 25 or less.

As used herein, a turbid liquid is one that is thick or opaque withvisible particles in suspension, for example, a liquid that is cloudy ormuddy in appearance.

As used herein, “clear” can be used to describe a composition asprovided herein, for example, the aqueous liquid dilution compositionscontaining the diluted nanoemulsion concentrates and/or the nanoemulsionconcentrates themselves. In one example, a clear liquid is one that doesnot appear cloudy by empirical observation (e.g., to the naked eye)and/or does not contain particles or crystals that are visible to thenaked eye, or that does not exhibit “ringing.” In another example, aclear liquid is one that has a low or relatively low turbidity value,for example an NTU value, that is less than or equal to a desired NTUvalue. In one example, a clear liquid has an NTU value of less than 300or less than about 300, typically less than 250 or less than about 250,typically less than 200 or less than about 200, for example, 200, 175,150, 100, 50, 25 or less. In another example, a liquid is clear if ithas a turbidity value (NTU) of 30 or about 30; or an NTU value of lessthan 30 or about 30, for example, less than 29 or about 29, less than 28or about 28, less than 27 or about 27, less than 26 or about 26, lessthan 25 or about 25, less than 24 or about 24, less than 23 or about 23,less than 22 or about 22, less than 21 or about 21, less than 20 orabout 20, less than 19 or about 19, less than 18 or about 18, less than17 or about 17, less than 16 or about 16, less than 15 or about 15, lessthan 14 or about 14, less than 13 or about 13, less than 12 or about 12,less than 11 or about 11, less than 10 or about 10, less than 9 or about9, less than 8 or about 8, less than 7 or about 7, less than 6 or about6, less than 5 or about 5, less than 4 or about 4, less than 3 or about3, less than 2 or about 2, less than 1 or about 1; or 29 or about 29, 28or about 28, 27 or about 27, 26 or about 26, 25 or about 25, 24 or about24, 23 or about 23, 22 or about 22, 21 or about 21, 20 or about 20, 19or about 19, 18 or about 18, 17 or about 17, 16 or about 16, 15 or about15, 14 or about 14, 13 or about 13, 12 or about 12, 11 or about 11, 10or about 10, 9 or about 9, 8 or about 8, 7 or about 7, 6 or about 6, 5or about 5, 4 or about 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0or about 0. In another example, a clear liquid is one that has a smallor relatively small average particle size (e.g., less than 1000 nm orabout 1000 nm, typically less than 500 nm or less than about 500 nm,typically less than 300 nm or about 300 nm, typically less than 250 nmor about 250 nm, typically less than 200 nm or about 200 nm, forexample, less than 150 or about 150 nm, less than 100 nm or about 100nm, less than 75 nm or about 75 nm, less than 50 nm or about 50 nm, lessthan 25 nm or about 25 nm or less than 10 nm or about 10 nm), forexample, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80,90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm.

In another example, clarity is expressed relatively. For example, it canbe desired that a particular composition is equally as clear, about asclear, or more clear than another liquid (as measured empirically, or bymeasuring turbidity value or particle size). For example, clarity can beassessed relative to another aqueous liquid dilution composition, forexample, a beverage. For example, In one example, a liquid is clear ifit is similar in appearance to another clear liquid, for example, abeverage, for example, water. For example, it can be desired that acomposition has a particle size that is less than or equal to anotherliquid, for example, a beverage. In another example, it can be desiredthat a composition has a turbidity value that is less than or equal toanother liquid, for example, a beverage. Thus, when the nanoemulsionconcentrates provided herein are diluted into an aqueous medium, theresulting compositions is about as clear or is as clear or is withinabout 10, 20, 30, 40, 50 NTU of the aqueous medium prior to addition ofthe concentrate.

As used herein, “hydrophilic” and “polar” refer synonymously toingredients and/or compounds having greater solubility in aqueousliquids, for example, water, than in fats, oils and/or organic solvents(e.g., methanol ethanol, ethyl ether, acetone and benzene).

Exemplary of the polar ingredients in the provided compositions arepolar solvents, which are solvents more readily miscible with water andother polar ingredients. Thus, polar ingredients are more readilydissolved in polar solvents than non-polar solvents. Polar solvents arewell-known. The polarity of a solvent can be assessed by measuring anumber of different parameters according to well known methods asdescribed herein (see, e.g., Prizbytek, “High Purity Solvent Guide,”Burdick and Jackson Laboratories, Inc., 1980). Polar solvents generallyhave high dielectric constants, typically dielectric constants greaterthan at or about 15, such as at or about 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60,65, 70, 75, 80 85, 90, or greater than 90, and generally have highpolarity indices, typically greater than at or about 3, such as at orabout 3, 4, 5, 6, 7, 8 or 9 or greater than 9. Polar solvents generallyhave large dipole moments, typically greater than at or about 1.4 Debye,such as at or about, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3,2.4, 2.5, 2.6, 3.0, 3.5, 4 or greater than 4 Debye. Polar solventsinclude polar protic solvents and polar aprotic solvents.

As used herein, a polar protic solvent is a polar solvent containing ahydrogen atom attached to an electronegative atom, such that thehydrogen has a proton-like character and/or the bond between thehydrogen and electronegative atom is polarized. Exemplary polar proticsolvents include, but are not limited to water, alcohols, includingmonohydric, dihydric and trihydric alcohols, including, but not limitedto, methanol, ethanol, glycerin, propylene glycol.

Dihydric alcohols are alcohols containing two hydroxyl groups. Exemplarydihydric alcohols include, but are not limited to, glycols, e.g.,propylene glycol, ethylene glycol, tetraethylene glycol, triethyleneglycol, trimethylene glycol.

Trihydric alcohols are alcohols containing three hydroxyl groups.Exemplary trihydric alcohols include, but are not limited to glycerin,butane-1,2,3-triol, pentane -1,3,5-triol,2-amino-2-hydroxymethyl-propane-1,3-diol.

Monohydric alcohols are alcohols containing a single hydroxyl group,including but not limited to, methanol, ethanol, propanol, isopropanol,n-butanol and t-butanol. In one example, the polar solvent in theprovided compositions is not a monohydric alcohol.

As used herein, “non-polar”, “lipophilic”, and “lipid-soluble”synonymously refer to compounds (e.g., non-polar compounds) and/oringredients, for example, non-polar active ingredients, which havegreater solubility in organic solvents (e.g., ethanol, methanol, ethylether, acetone, and benzene) and in fats and oils, than in aqueousliquids, for example, water. Non-polar compounds include drugs,hormones, vitamins, nutrients and other lipophilic compounds. Typically,the non-polar compounds used in the provided compositions are poorlywater soluble, for example, water insoluble or compounds having lowwater solubility. Exemplary non-polar compounds include non-polar activeingredients, for example, lipid-soluble drugs, hormones, essential fattyacids, for example, polyunsaturated fatty acids (PUFA), for example,omega-3 and omega-6 fatty acids, vitamins, nutrients, nutraceuticals,minerals and other compounds. Additional exemplary non-polar compoundsare described herein. The provided compositions can be formulated withany non-polar compound, for example, non-polar active ingredient.

As used herein, non-polar active ingredient refers to a non-polarcompound that, when administered to a subject, for example, a human,induces or is proposed to induce a desired response, such as alteringbody function at the cellular, tissue, organ or other level, and/oraltering cosmetic appearance or other property, or a non-polar compoundthat is ingested in order to achieve a desired effect. Non-polar activeingredients can be any synthetic or natural non-polar ingredient orcompound, including a pharmaceutical, drug, therapeutic, nutritionalsupplement, herb, hormone or other ingredient. Non-polar activeingredients can include the non-polar active ingredients listed herein,as well as other pharmaceutically acceptable or food-grade activederivatives of the active ingredients, for example, salts, esters,amides, prodrugs, active metabolites, isomers, fragments and analogs.Active ingredients can include compounds proven to have a desired effectand also compounds thought to produce such effects, for example,compounds typically ingested for nutritional supplementation purposes.

As used herein, a subject includes an animal, typically a mammal,typically a human.

As used herein, an additive includes anything that one can add to afood, beverage, or other human consumable, to enhance one or more of itsnutritional, pharmaceutical, dietary, health, nutraceutical, healthbenefit, energy-providing, treating, holistic, or other properties. Forexample, provided herein are compositions and methods for preparingfoods, beverages and other aqueous human consumables, that include oneor more additives, typically oil based additives (e.g., non-polarcompounds), such as nutraceuticals, pharmaceuticals, vitamins, typicallyoil soluble vitamins, for example, Vitamin D, particularly D3, VitaminE, and Vitamin A, minerals, fatty acids, such as essential fatty acids,e.g., polyunsaturated fatty acids, for example, omega-3 fatty acids, andomega-6 fatty acids, for example, alpha-linolenic acid (ALA),docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), gamma-linolenicacid GLA, CLA, saw palmetto extract, flaxseed oil, fish oil, algae oil,phytosterols, and Coenzymes, for example, Coenzyme Q10 and otheradditives.

As used herein, an effective amount of an additive, such as a non-polarcompound, such as a non-polar active ingredient, refers to the quantityand/or concentration of the additive necessary for preventing, curing,ameliorating, arresting or partially arresting a symptom of a disease ordisorder, or the quantity and/or concentration desired by an individualfor intake, such as daily intake, and/or nutritional supplementation,for example, an amount sufficient to enhance the nutritional,pharmaceutical, nutraceutical, health or energy property of a food,beverage, or other consumable. In some examples, it is desired that theprovided compositions, for example, the liquid nanoemulsion concentratesand/or the liquid dilution compositions, contain an effective amount ofa particular non-polar compound, for example, a particular amount pervolume or weight of the composition.

In one example, an effective amount is a concentration or amount of aliquid nanoemulsion composition where at least 25 mg or about 25 mg,typically at least 35 mg, for example, 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190,200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 325, 350, 375,400, 425, 450, 475, 500, 550, 600, 700, 800, 900, 1000, 1500, 2000 mg,or more, of the non-polar active ingredient, is contained in at least 8fluid ounces of an aqueous medium, e.g., a beverage.

As used herein, unit dose form refers to physically discrete unitssuitable for human and animal subjects and packaged individually as isknown in the art.

As used herein, “water insoluble” refers to a property of a compound,none of which dissolves when the compound is mixed with water, forexample, when mixed with water at room temperature, for example, between25 and 50° C. or between about 25 and 50° C. In one example, thenon-polar compounds are water insoluble. In another example, thenon-polar compounds in the provided compositions are slightly soluble inwater, for example, having low water solubility.

As used herein, low water solubility refers water solubility of lessthan 30 or about 30 mg/mL, typically less than 20 mg/mL or about 20mg/mL, typically, less than 10 mg/mL or about 10 mg/mL, typically lessthan 1 mg/mL or about 1 mg/mL, for example, solubility in water of 30,29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12,11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 mg/mL or less, for example, when mixedwith water at room temperature, for example, between 25° C. and 50° C.or between about 25° C. and 50° C. As used herein, poorly water solublecan be used to refer to compounds, for example, non-polar compounds thatare water insoluble or have low water solubility.

As used herein, concentrate, liquid concentrate and liquid nanoemulsionconcentrate, are used synonymously to refer to provided compositionsthat contain the non-polar compounds, are liquid at room temperature,for example at 25° C. or about 25° C., or at a temperature of between25° C. or about 25° C. and 50° C. or about 50° C., and can be diluted inaqueous media to form the provided aqueous liquid dilution compositions.Typically, the liquid nanoemulsion concentrate is an emulsionconcentrate that has a particle (droplet) size (or can be diluted toform an aqueous liquid dilution composition having a particle size) thatis less than 1000 or about 1000, typically less than 500 or about 500,typically less than 300 or about 300 nm, typically less than 250 orabout 250 nm, for example, less than 200 or about 200, for example, lessthan 150 or about 150 nm, for example, a particle size equal to, lessthan or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100,110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm.

As used herein, liquid composition is used to refer to any liquid, forexample, a composition that is a liquid at room temperature, forexample, at 25° C. or about 25° C., or at a temperature of between 25°C. or about 25° C. and 50° C. or about 50° C. Exemplary of the providedliquid dilution compositions are aqueous liquid dilution compositionsinto which one or more liquid nanoemulsion concentrate has been diluted,for example, aqueous liquid dilution compositions containing the dilutedconcentrates. In this example, the non-polar compound and otherlipophilic compounds in the concentrate form the dispersion phase withinthe aqueous liquid, which is an emulsion (e.g., nanoemulsion).

As used herein, “liquid dilution composition” “dilution composition” and“liquid dilution” are used synonymously to refer to a composition thatcontains one or more of the provided liquid nanoemulsion concentrates(e.g., the liquid nanoemulsion concentrates containing the non-polarcompound(s)), diluted in a liquid, for example, an aqueous medium.Exemplary of the provided liquid dilution compositions are aqueousliquid dilution compositions, for example, beverages or other liquidscontaining the liquid nanoemulsion concentrates, for example, water,sauces, soups, syrups, soda, juice, for example, fruit juice, milk,coffee, tea, nutritional beverages, sports drinks, energy drinks,vitamin-fortified beverages, flavored water, and other beveragescontaining the diluted concentrates.

As used herein, aqueous liquid dilution compositions are liquid dilutioncompositions that are primarily aqueous, for example, a compositioncomprising a liquid nanoemulsion concentrate diluted in an aqueousmedium, for example, water or other beverage. It is not necessary thatthe aqueous liquid dilution composition is completely aqueous. Forexample, the aqueous liquid dilution compositions can contain an aqueousportion, for example, an aqueous continuous phase, as well as anadditional portion, for example, a dispersion phase, for example, alipophilic dispersion phase. Typically, the lipophilic dispersion phasecontains one or more lipophilic substances, for example, one or morenon-polar compounds, for example, non-polar active ingredients.Exemplary of the provided aqueous liquid dilution compositions arebeverages containing the active ingredients, for example, water, soda,juice, for example, fruit juice, milk, coffee, tea, nutritionalbeverages, sports drinks, energy drinks, vitamin-fortified beverages,flavored water, and other beverages. Typically, the aqueous liquiddilution compositions are beverages including the non-polar compound,for example, beverages containing the diluted concentrates.

As used herein, “oil phase” is used to refer to the portion (or phase)of a composition such as those provided herein that contains one or morelipophilic ingredients and/or amphiphilic ingredients (oil phaseingredients) and is, in general, the lipid-soluble phase. In theprovided emulsion compositions (e.g., the nanoemulsion concentrates andthe dilution compositions), the oil phase typically represents thedispersion phase. “Oil phase” also can be used to refer to the liquidcontaining the oil phase ingredients that is generated, typically in anoil phase vessel, while carrying out the methods for making the liquidnanoemulsion concentrates. For example, oil phase can refer to themixture of the components (oil phase ingredients) that are combined,mixed and heated, for example, in the oil phase vessel (e.g., tank),prior to mixing with the water phase. “Oil phase” can refer to the oilphase mixture that is formed after all the ingredients are dissolved;alternatively, it can refer to the forming mixture, for example, as itis being mixed/heated.

As used herein, oil phase ingredient(s) refers to the components of theprovided compositions that are included in the oil phase in the providedmethods for making the compositions. Typical oil phase ingredientsinclude non-polar compounds, e.g., non-polar active ingredients;surfactants; co-surfactants; oils, such as non-polar solvents;preservatives; and emulsion stabilizers. Other lipophilic and/oramphiphilic ingredients can be included in the oil phase.

As used herein, “water phase” is used to refer to the portion (phase) ofa compositions such as those provided herein that contains one or morehydrophilic ingredients and/or amphiphilic ingredients (water phaseingredients) and is, in general, the water-soluble phase. Typically, inthe provided emulsion compositions, for example, the nanoemulsionconcentrates and the dilution compositions, the water phase is thecontinuous phase. “Water phase” also is used to refer to the liquidcontaining the water phase ingredients that is generated while carryingout the methods for making the liquid nanoemulsion concentrates. Forexample, water phase can refer to the mixture of the components (waterphase ingredients) that are combined, mixed and heated, for example, inthe water phase tank, prior to mixing with the oil phase. “Water phase”can refer to the water phase mixture that is formed after all theingredients are dissolved; alternatively “water phase” can refer to theforming mixture, for example, as it is being mixed/heated.

As used herein, water phase ingredient(s) refers to the components ofthe provided compositions that are included in the water phase (e.g.,added to the water phase vessel) in the provided methods for making thecompositions. Typical water phase ingredients include, but are notlimited to, polar solvents, typically polar protic solvents, such aswater and alcohols, typically alcohols having more than one hydroxygroup such as dihydroxy and trihydroxy alcohols, e.g., glycerol andpropylene glycol; surfactants; co-surfactants; preservatives; andemulsion stabilizers. Other hydrophilic and/or amphiphilic ingredientscan be included in the water phase.

As used herein, an initial concentrate is a concentrate (e.g., liquidnanoemulsion concentrate) that is made in the provided methods offormulating the provided concentrates, typically by selectingingredients, for example, surfactant(s), non-polar compound(s), polarsolvent, and, optionally, other ingredients, and selecting startingconcentrations of the ingredients from an appropriate concentrationrange as described herein.

As used herein, stability refers to a desirable property of the providedcompositions, for example, the ability of the provided compositions toremain free from one or more changes over a period of time, for example,at least or over 1, 2, 3, 4, 5, 6 or more days, at least or over 1, 2,3, 4, or more weeks, at least or over 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12 or more months, or at least or over 1, 2, 3, 4 or more years. In oneexample, the composition is stable if it is formulated such that itremains free from oxidation or substantial oxidation over time. Inanother example, the stable compositions remain clear over time. Inanother example, the stable compositions remain safe and/or desirablefor human consumption over time. In one example, stability refers to thelack of precipitates forming in the compositions over the period oftime. In a related example, stability refers to the lack of “ringing”over the period of time. In another example, the composition is stableif it does not exhibit any visible phase separation over a period oftime, for example, after 6, 12, 24 hours, two, three, four, five days,after one week or after one month and longer, including up to 6 months.In one example, the compositions are stable if they exhibit one or moreof these described characteristics, over time, when kept at a particulartemperature. In one example, the compositions remain stable at roomtemperature, for example, 25° C. or about 25° C. In another example, thecompositions remain stable at between 19° C. and 25° C. In anotherexample, the compositions remain stable at refrigerated temperatures,for example, 4° C. or about 4° C., or at frozen temperature, forexample, at −20° C. or about −20° C.

As used herein, stabilize means to increase the stability of one of theprovided compositions.

As used herein, room temperature and ambient temperature are used todescribe a temperature that is common in one or more enclosed spaces inwhich human beings typically are or reside. Room temperature can vary,but generally refers to temperatures between 19° C. or about 19° C. and25° C. or about 25° C. When a composition is stored at room temperature,it should be understood it is generally kept at a temperature withinthis range or about within this range.

As used herein, refrigerated temperature refers to a temperature that iscommon in a refrigerator, for example, a household or restaurantrefrigerator, for example, a temperature that is cooler than roomtemperature, but typically a few degrees above the freezing point ofwater (0°C.). Typically, refrigerated temperatures are between about 10°C. or about 10° C. an 0° C. or about 0° C., for example, 4° C. or about4° C. When a composition is stored at a refrigerated temperature, itshould be understood that it is kept at a temperature common tohousehold or industrial refrigerators.

As used herein, frozen temperature refers to a temperature around orbelow the freezing point of water, e.g., a temperature commonly used ina household freezer, for example, 0° F. or about 0° F., for example,−19° C. or about −19° C. or −20° C. or about −20° C., or colder.

As used herein, ranges and amounts can be expressed as “about” aparticular value or range. About also includes the exact amount. Hence“about 5 grams” means “about 5 grams” and also “5 grams.’ It also isunderstood that ranges expressed herein include whole numbers within theranges and fractions thereof. For example, a range of between 5 gramsand 20 grams includes whole number values such as 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19 and 20 grams, and fractions within therange, for example, but not limited to, 5.25, 6.72, 8.5, and 11.95grams.

As used herein, “optional” or “optionally” means that the subsequentlydescribed event or circumstance does or does not occur and that thedescription includes instances where said event or circumstance occursand instances where it does not. For example, an optionally variantportion means that the portion is variant or non-variant. In anotherexample, an optional ligation step means that the process includes aligation step or it does not include a ligation step.

As used herein, “ringing” refers to the formation of a whitish or opaquering around a container containing a liquid, for example, an aqueousliquid, for example a beverage, for example, a liquid dilutioncomposition containing an emulsion or nanoemulsion. Typically, the ringforms around the perimeter of the container, typically at the surfacelevel of the liquid in the container, for example, at the neck of thecontainer. Ringing can occur over time and, if it occurs over a shortperiod of time, can be a sign of instability. Ringing typically isundesirable, particularly in the case of a liquid for human consumption,for example, a beverage. Typically, the provided compositions do notexhibit “ringing” or are stable, without ringing, for a long period oftime, for example, days, weeks, months or years. In one example, thecompositions are free from ringing over time, when kept, for example, atroom temperature, refrigerated and/or frozen.

As used herein, fatty acid refers to straight-chain hydrocarbonmolecules with a carboxyl (COOH) group at one end of the chain.

As used herein, polyunsaturated fatty acid and PUFA are usedsynonymously to refer to fatty acids that contain more than onecarbon-carbon double bond in the carbon chain of the fatty acid. PUFAs,particularly essential fatty acids, are useful as dietary supplements.

As used herein, essential fatty acids are PUFAs that mammals, includinghumans, cannot synthesize using any known chemical pathway. Thus,essential fatty acids must be obtained from diet or by supplementation.Exemplary of essential PUFA fatty acids are omega-3 (ω3; n-3) fattyacids and the omega-6 (ω6; n-6) fatty acids.

As used herein, omega-3 (ω3; n-3) fatty acid are methylene interruptedpolyenes, which have two or more cis double bonds, separated by a singlemethylene group and in which the first double bond appears at the thirdcarbon from the last (ω) carbon. Omega-3 fatty acids are used as dietarysupplements, for example, for disease treatment and prevention. In oneexample, the provided compositions contain non-polar active ingredientsthat contain at least one omega-3 fatty acids. Exemplary of Omega-3fatty acids are Alpha-Linolenic acid (α-Linolenic acid; ALA) (18:3ω3) (ashort-chain fatty acid); Stearidonic acid (18:4ω3) (a short-chain fattyacid); Eicosapentaenoic acid (EPA) (20:5ω3); Docosahexaenoic acid (DHA)(22:6ω3); Eicosatetraenoic acid (24:4ω3); Docosapentaenoic acid (DPA,Clupanodonic acid) (22:5ω3); 16:3 ω3; 24:5 ω3 and nisinic acid (24:6ω3).Longer chain Omega-3 fatty acids can be synthesized from ALA (theshort-chain omega-3 fatty acid). Exemplary of non-polar activeingredients containing omega-3 fatty acids are non-polar activeingredients containing DHA and/or EPA, for example, containing fish oil,krill oil and/or algae oil, for example, microalgae oil, non-polaractive ingredients containing alpha-linolenic acid (ALA), for example,containing flaxseed oil.

As used herein, omega-6 (ω-6; n-6) fatty acids are methylene interruptedpolyenes, which have two or more cis double bonds, separated by a singlemethylene group and in which the first double bond appears at the sixthcarbon from the last (ω) carbon. In one example, the providedcompositions contain non-polar active ingredients that contain at leastone omega-3 fatty acids. Exemplary of Omega-6 fatty acids are Linoleicacid (18:2ω6) (a short-chain fatty acid); Gamma-linolenic acid (GLA)(18:3ω6); Dihomo gamma linolenic acid (DGLA) (20:3ω6); Eicosadienoicacid (20:2ω6); Arachidonic acid (AA) (20:4ω6); Docosadienoic acid(22:2ω6); Adrenic acid (22:4ω6); and Docosapentaenoic acid (22:5ω6).Exemplary of non-polar active ingredients containing omega-6 fatty acidsare ingredients containing GLA, for example, borage oil. Also exemplaryof PUFA-containing non-polar active ingredients are compounds containingconjugated fatty acids, for example, Conjugated linoleic acid (CLA) andcompounds containing saw palmetto extract.

As used herein, algae oil refers to any oil derived from marinedinoflagellates in, for example, microalgae, for example,Crypthecodinium sp, particularly, Crypthecodinium cohnii. In oneexample, algae oil is used as a non-polar compound, for example, as anactive ingredient, in the provided compositions. The algae oil typicallycontains DHA. In one example, the algae oil is also a source of EPA.

As used herein, fish oil refers to any oil derived from any fish,typically a cold water fish, for example, from fish tissue, for example,from frozen fish tissue, for example, from cod liver. In one example,fish oil is used as a non-polar compound, for example, an activeingredient, in the provided compositions. The fish oil typicallycontains DHA. In one example, the fish oil also contains EPA.

As used herein, preservative and preservatives are used synonymously torefer to ingredients that can improve stability of the providedcompositions. Preservatives, particularly food and beveragepreservatives, are well known. Any known preservative can be used in theprovided compositions. Exemplary of the preservatives that can be usedin the provided compositions are oil soluble preservatives, for example,benzyl alcohol, Benzyl Benzoate, Methyl Paraben, Propyl Paraben,antioxidants, for example, Vitamin E, Vitamin A Palmitate and BetaCarotene. Typically, a preservative is selected that is safe for humanconsumption, for example, in foods and beverages, for example, a GRAScertified and/or Kosher-certified preservative, for example, benzylalcohol.

As used herein, a solvent is ingredient that can be used to dissolveanother ingredient. For example, the solvents include polar andnon-polar solvents. The non-polar solvents include oils and othernon-polar ingredients that dissolve non-polar compounds. In one example,the non-polar active ingredient is dissolved in a non-polar solvent inpracticing the methods of producing the provided compositions. In thisexample, the provided compositions contain non-polar solvents in amountssufficient to dissolve the non-polar active ingredient. More than onenon-polar solvent can be used. Typically, the non-polar solvent is anoil that is included in the composition in addition to the non-polarcompound. For example, the non-polar solvent typically is not thenon-polar compound itself, e.g., is distinct from the non-polar solvent.Certain compounds, for example, flaxseed oil and safflower oil, can bnon-polar solvents and non-polar active ingredients. Typically, thenon-polar solvent contains one or more oils, typically oils other thanthe non-polar active ingredient or oil(s) not contained in the activeingredient. Exemplary non-polar solvents include, but are not limitedto, oils (in addition to the non-polar active ingredient), for example,Vitamin E oil, flaxseed oil, CLA, Borage Oil, D-limonene, Canola oil,corn oil, MCT oil and oat oil. Other oils also can be used. Exemplary ofthe Vitamin E oil is the oil sold by ADM Natural Health and Nutrition,Decatur, IL, under the name Novatol™5-67 Vitamin E (D-alpha-Tocopherol;ADM product code 410217). This Vitamin E oil contains at least 67.2%Tocopherol and approximately 32.8% soybean oil. In one example, thenon-polar solvent is referred to, synonymously as “non-polarsolubilizer.”

As used herein, “w/w,” “weight per weight,” “by weight” “% by weight”and “weight percent” are used synonymously used to express the ratio ofthe mass of one component of a composition compared to the mass of theentire composition. For example, when the amount of a particularingredient represents 1%, by weight (w/w) of a concentrate, the mass ofthat ingredient is 1% of the mass of the entire concentrate. Similarly,when the amount of an ingredient is 50% (w/w) of the concentrate, themass of that ingredient is 50% of the entire mass of the concentrate.Similarly, when a composition and/or a compound contains 10%, by weightof an ingredient, the mass of the ingredient is 10% of the total mass ofthe composition or compound. When only a concentration, amount, orpercentage (without units) is listed, it is to be understood that theconcentration or percentage is a concentration or percentage, by weight.

Similarly, as used herein “v/v,” “volume per volume,” “percent byvolume” and “volume percent” are used synonymously to express the ratioof the volume of one component of a composition and the volume of theentire composition.

As used herein, emulsion stabilizer refers to compounds that can be usedto stabilize and/or emulsify and/or change the viscosity of the providedcompositions, for example, the liquid nanoemulsion concentrate and/orthe aqueous compositions containing the diluted concentrates. In oneexample, the emulsion stabilizer increases the viscosity of the liquidconcentrate. In one example, one or more emulsion stabilizers is added,during formulation, after evaluation of an initial concentrate,particularly if the oil and water phases of the initial concentrate (orthe aqueous liquid dilution composition resulting from dilution of theinitial concentrate) appear to be separating. Addition of the emulsionstabilizer can prevent separation of the oil and water phases.

Exemplary of an emulsion stabilizer that can be used in the providedcompositions is a composition containing a blend of gums, for example,gums used as emulsifying agents, for example, a blend containing one ormore of xanthan gum, guar gum and sodium alginate, for example, theemulsion stabilizer sold under the brand name SALADIZER®, available fromTIC Gums, Inc. (Belcamp, Md.). Other gums can be included in theemulsion stabilizer, for example, gum acacia and sugar beet pectin.Other blends of similar gums can also be used as emulsion stabilizers.

As used herein, a pH adjuster is any compound, typically an acid or abase, that is capable of changing the pH of the provided compositions,for example, to reduce the pH of the composition or to increase the pHof the composition, typically without altering other properties of thecomposition, or without substantially altering other properties. pHadjusters are well known. Exemplary of the pH adjusters are acids, forexample, citric acid and phosphoric acid, and bases.

As used herein, flavor is any ingredient that changes, typicallyimproves, the taste and/or smell of the provided composition, forexample, the aqueous liquid dilution compositions, for example, thebeverages.

As used herein, “not more than” and “NMT” refer to a quantity that isless than or equal to the listed quantity. Similarly, “not less than”and “NLT” refer to a quantity that is greater than or equal to thelisted quantity.

As used herein, natural is used to refer to a composition, and/oringredients in the composition, that can be found in nature and is notsolely man-made. For example, benzyl alcohol is a natural preservative.In one example, the natural composition/ingredient is GRAS and/orKosher-certified. Typically, the provided compositions are natural,semi-natural and/or contain one or more natural ingredients.

As used herein, “G.R.A.S.” and “GRAS” are used synonymously to refer tocompounds, compositions and ingredients that are “Generally Regarded asSafe” by the USDA, FDA for use as additives, for example, in foods,beverages and/or other substance for human consumption, for example, anysubstance that meets the criteria of sections 201(s) and 409 of the U.S.Federal Food, Drug and Cosmetic Act. Typically, the compositionsprovided herein are GRAS certified.

As used herein, kosher is used to refer to substances that conform toJewish Kosher dietary laws, for example, substances that do not containingredients derived from non-kosher animals or ingredients that were notmade following kosher procedures. Typically, the compositions providedherein are Kosher certified.

As used herein, vessel refers to any container, for example, tanks,pots, vials, flasks, cylinders and beakers, that can be used to containthe ingredients and/or phases of the provided compositions, during themethods for making the compositions. In one example (e.g., for theprovided scaled-up methods), the vessel is a tank, which is used to mixand/or heat one or more ingredients and/or phases of the compositions,for example, water phase tanks and oil phase tanks. Typically, the oiland the water phases are mixed and heated in separate tanks, beforecombining the phases to form an emulsion. In another example, the tankis a packaging or holding tank, which holds the provided compositionsafter forming the compositions, for example, the emulsions. A number oftanks are available for mixing ingredients. Typically, the tanks arecleaned, for example, rinsed, soaped and/or sanitized according to knownprocedures, prior to use and between uses. Typically, the tanks areequipped with one or more mixers, for example, a standard mixer and/orhomogenizer, which are used to mix the ingredients added to the tank. Inone example, the tank further is equipped with a heating and/or coolingdevice. For example, the tank can be a water-jacketed tank. Thetemperature of the water-jacketed tank is controlled through thewater-jacket, for example, to heat the contents, for example, whilemixing.

As used herein, a water phase vessel refers to the vessel used to mixand/or heat the water phase ingredients to generate the water phase ofthe provided compositions. In one example (e.g., for the scaled-upmethods), the water phase vessel is a water phase tank. In one example,the water phase tank is a water jacketed tank, which is equipped with awater jacket that can be used to heat the contents of the tank.

As used herein, an oil phase vessel refers to the vessel used to mixand/or heat the oil phase ingredients to generate the oil phase of theprovided compositions. Typically, the oil phase vessel is an oil phasetank. In one example, the oil phase tank is a water jacketed tank.

As used herein, transfer means refers to any equipment, combination ofequipment and/or system that can be used to transfer liquid, forexample, from one tank to another tank, in the provided methods formaking the compositions. Exemplary of the transfer means are a transferpump and appropriate fittings, for example, sanitary fittings, ballvalves and transfer hoses, for example, food grade hoses.

As used herein, a mixer is any piece of equipment or combination ofequipment that can be used to mix ingredients in the provided methodsfor making the compositions, for example, standard mixers andhomogenizers (shears). For example, mixers can be used to mix theingredients of the water phase, the oil phase, and/or to mix theadditional ingredients.

As used herein, standard mixers are mixers that are used to combine agroup of ingredients, for example, the oil phase ingredients or thewater phase ingredients, or to mix one or more ingredients with aliquid, for example, with an emulsion, for example, to mix additionalingredients with the emulsion. Standard mixers can be any mixers thatmove the material, for example, the ingredients, during heating, forexample, to promote dissolving of the ingredients.

As used herein, “homogenizer” and “shear” are used to refer to mixersthat typically have high shear, which can be used, for example, to forman emulsion, for example, to emulsify the water phase and the oil phase,in the provided methods. The homogenizers typically are capable ofhigh-shear mixing, which emulsifies the phases.

As used herein, a cooling apparatus is any piece of equipment orcombination of equipment that can be used with the provided methods tocool the compositions and phases and ingredients thereof, for example,during mixing and/or homogenizing, for example, to chill the mixturewhile emulsifying the oil and water phases. Exemplary of the coolingapparatuses are coolers (chillers), for example, recirculating coolerswhich can be attached, for example, to the tanks used in the providedmethods, for example, remotely or by a tank mounted in the cooler, torecirculate fluid from the tank, through the chiller and back to thetank, in order to rapidly cool and maintain the temperature of themixture during mixing. Typically, the cooling apparatus can be used tocool the liquid to between 25° C. or about 25° C. and 45° C. or about45° C., for example, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44 or 45° C., typically between 25° C. and 43°C., typically between 35° C. and 43° C., for example, 26.5° C.

As used herein, rapid cooling refers to a process by which acomposition, for example, a liquid composition, for example, a formingemulsion, is cooled to a desired temperature, for example, between 25°C. or about 25° C. and 45° C. or about 45° C., typically between 35° C.and 43° C., for example, 26.5° C., in less than 2 hours or about 2hours, typically less than 1 hour or about 1 hour, for example, in atleast between 30 minutes or about 30 minutes and 60 minutes or about 60minutes, for example, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59or 60 minutes.

As used herein, low heat refers to a temperature between 45° C. or about45° C. and 85° C. or about 85° C., for example, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85°C., for example, not more than 85° C. or about 85° C., typically notmore than 60° C. or about 60° C., typically, 60° C. or 60° C. In theprovided methods for making the liquid nanoemulsion concentrates, theoil phase and water phase ingredients typically are heated, using lowheat, in order to preserve the ingredients, for example, in order toprevent oxidation of the ingredients, for example, the non-polar activeingredients, for example, the omega-3 containing compounds, for example,the DHA.

As used herein, “consisting essentially of,” means containing thefollowing list of ingredient(s), and not including any additional activeingredient, for example, not including any additional active drug orpharmaceutical. For example, a composition, for example, a liquidnanoemulsion, consisting essentially of a listed plurality ofingredients contains those particular ingredients and does not containany additional active drug or pharmaceutical.

As used herein, C₁-C_(x) includes C₁-C₂, C₁-C₃ . . . C_(x).

As used herein, a sucrose fatty acid ester is a compound having theformula shown in Scheme V, below.

where each of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ independently is:

a hydroxyl (—OH) group, or

where:

each R is an alkyl group having 3-27 carbon atoms; and

when more than one of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ is

each R can be a different alkyl group (e.g., having different number ofcarbon atoms and/or different saturation), or can be the same alkylgroup.

In some examples, the provided compositions contain sucrose fatty acidester surfactants that have sucrose fatty acid ester(s) havingstructures according to Scheme V above, where R typically containsbetween 7 and 27 carbon atoms. Typically, the sucrose fatty acid estersurfactants contain sucrose fatty acid monoesters. The sucrose fattyacid ester surfactants include mixtures, or blends, of sucrose fattyacid esters, which typically include monoesters, and can also includediesters, triesters and polyesters.

A sucrose fatty acid monoester has the structure set forth in Scheme V,where one of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ (typically X¹) is:

and the other seven of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ are each,independently, —OH.

An exemplary sucrose fatty acid monoester has the following structure:

where R is an alkyl group having 3-27 carbons.

Sucrose fatty acid diesters, sucrose fatty acid triesters, and sucrosefatty acid polyesters, respectively, are sucrose fatty acid estershaving structures according to Scheme V, above, where two (diesters),three (triesters) or more (polyesters) of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ andX⁸, (and typically X¹ and X⁸) independently, are

Thus, the sucrose fatty acid ester surfactants include sucrosemonoesters, diesters, triesters and polyesters, and mixtures thereof,and typically contain sucrose monoesters. The sucrose fatty acid estersurfactants include single fatty acid esters and also includehomogeneous mixtures of sucrose esters, containing two or more sucrosefatty acid esters with different lengths of fatty acid carbon chainsand/or members with different degrees of esterification. For example,the sucrose fatty acid ester surfactants include mixtures of monoesters,diesters, triesters, and/or polyesters. The sucrose fatty acid estermixtures can include one or more sucrose fatty acid esters, such as, butnot limited to, sucrose stearate, sucrose laurate, sucrose palmitate,sucrose oleate, sucrose caprylate, sucrose decanoate, sucrose myristate,sucrose pelargonate, sucrose undecanoate, sucrose tridecanoate, sucrosepentadeconoate and sucrose heptadecanoate, and homologs thereof.

As used herein, the term “alkyl” and “alkyl group” refer to straight orbranched chain substituted or unsubstituted hydrocarbon groups havingany number of carbon atoms; number of carbon atoms can be specified, forexample, 1 to 30 carbon atoms, 8 to 28 carbon atoms, 7 to 27 carbonatoms, 8 to 22 carbon atoms, 8 to 20 carbon atoms, 8 to 18 carbon atomsand 12 to 18 carbon atoms. An alkyl group can be a “saturated alkyl,”meaning that it does not contain any alkene or alkyne groups or an“unsaturated alkyl,” meaning that it contains at least one alkene oralkyne group, an optionally can be substituted. An alkyl group thatincludes at least one carbon-carbon double bond (C═C) also is referredto by the term “alkenyl;” alkenyl groups optionally can be substituted.An alkyl group that includes at least one carbon-carbon triple bond(C≡C) also is referred to by the term “alkynyl;” alkynyl groupsoptionally can be substituted.

As used herein, the singular forms “a,” “an” and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to a compound comprising “an extracellular domain”includes compounds with one or a plurality of extracellular domains.

B. Compositions Containing Non-Polar Compounds

Provided herein are compositions containing non-polar compounds andmethods for making the compositions. Non-polar compounds are poorlywater soluble (e.g., having low water solubility or beingwater-insoluble). Thus, it generally can be difficult to formulatenon-polar compounds into compositions for human consumption,particularly aqueous compositions, for example, foods and beverages.Poor water solubility of non-polar compounds also can contribute totheir poor bioavailability. Improved methods and compositions forformulating non-polar compounds are provided herein.

In general, emulsions (e.g., oil-in-water emulsions) are colloidaldispersions of two immiscible liquids (e.g., oil and water or otheraqueous liquid), containing a continuous and a dispersed phase.Emulsions can be used to disperse non-polar compounds in aqueousliquids. In an oil-in-water emulsion, the dispersed phase is an oilphase and the continuous phase is an aqueous (water) phase. There is aneed for emulsions (e.g., oil-in-water emulsions) containing non-polarcompounds in aqueous liquids and methods and compositions for generatingthe dilutions compositions, such as beverages, that are clear andstable. In particular, emulsions are needed that are more suitable anddesirable for human consumption of the non-polar compounds, for example,in foods and beverages. For example, emulsions having improved clarity(e.g., small particle size, low turbidity), stability (e.g., lack ofseparation), taste and smell, particularly when diluted into a beverageto provide a desired dosage of an active ingredient are needed and areprovided herein.

Among the provided compositions are such improved emulsions. Forexample, emulsions are provided that contain the non-polar compoundsdispersed in aqueous liquid and have desirable properties, includingimproved clarity, stability, smell and taste. The provided compositions(and methods for making the compositions) can be used to formulate anynon-polar compound in aqueous compositions, including the non-polarcompounds (e.g., non-polar active ingredients) described herein andother known non-polar compounds.

Typically, the provided emulsions containing the non-polar compounds arenanoemulsions, which are emulsions having dispersed droplets (particles)with diameters less than 1000 nm or less than about 1000 nm, typically,less than 500 nm or less than about 500 nm, typically less than 300 nmor about 300 nm, typically less than 250 or less than about 250 nm,typically less than 200 nm or less than about 200 nm, for example, lessthan or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100,110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm. Typically, theprovided nanoemulsion compositions are oil-in-water nanoemulsions,containing the non-polar compounds dispersed in aqueous liquid.

The provided emulsion compositions are stabilized by one or moresurfactants and/or co-surfactants and/or emulsion stabilizers.Surfactants form an interfacial film in the emulsion, between the oiland water phase, providing stability. Typically, the nanoemulsions ofthe provided compositions contain micelles, in which one or moresurfactant surrounds the non-polar active compound. The micelles aredispersed in the water phase.

The provided emulsion compositions include liquid nanoemulsionconcentrates containing the non-polar compounds, which can be diluted toprovide non-polar compounds in aqueous compositions, such as beverages.The liquid nanoemulsion concentrates can be diluted into a medium, forexample, an aqueous medium for example, a beverage, to form a liquiddilution composition (e.g., aqueous liquid dilution composition)containing the non-polar compound. Also exemplary of the providedcompositions are the liquid dilution compositions (e.g., aqueous liquiddilution compositions, which can be clear) made by diluting the liquidnanoemulsion concentrates in the medium.

The compositions can be made using any non-polar compound. The non-polarcompounds typically are non-polar active ingredients, for example,pharmaceuticals, nutraceuticals, vitamins and minerals. The non-polaractive ingredients include, but are not limited to, PolyunsaturatedFatty Acids (PUFA)-containing compounds, for example, omega-3-containingactive ingredients, for example, compounds containing ALA, DHA and/orEPA, for example, oils derived from fish and microalgae, krill and/orflaxseed extract, and omega-6-containing non-polar active ingredients,for example, gamma-linolenic acid (GLA)-containing compounds, forexample, borage oil; saw palmetto oil-containing compounds; conjugatedfatty acid containing-ingredients, for example, Conjugated Linoleic acid(CLA)-containing compounds; coenzyme Q-containing active ingredients,for example, Coenzyme Q10 (CoQ10), typically oxidized CoQ10(ubidecarenone)-containing compounds; and compounds containingphytosterols (plant sterols). Additional exemplary non-polar activeingredients are described herein. Any non-polar compound can be used inthe provided compositions.

1. Liquid Nanoemulsion Concentrates Containing the Non-Polar Compounds

Provided are liquid nanoemulsion concentrates (also called“concentrates” or “liquid concentrates”) containing one or morenon-polar compounds. The concentrates can be diluted into aqueous mediato form aqueous liquid dilution compositions containing the non-polarcompounds. The liquid concentrates are formulated based on one or moredesirable properties, for example, clarity; safety; taste; smell;stability, for example, lack of phase separation, “ringing” and/orprecipitation over time, and/or bioavailability of the concentrateand/or the aqueous liquid dilution compositions containing theconcentrate. In one example, the desirable property is the ability ofthe provided concentrate to yield a clear or partially clear aqueousliquid dilution composition when it is diluted into aqueous medium, forexample, a beverage such as water. In another example, the desirableproperty relates to the safety of the concentrates and/or thedesirability of the concentrates for human consumption, for example, infoods and beverages. In another example, it can be desirable that theconcentrate contains less than or equal to a particular concentration ofone or more ingredients. In another example, it can be desirable thatthe concentrate contains greater than or equal to a particularconcentration of one or more ingredients.

In addition to the non-polar compounds, the concentrates contain atleast one surfactant. Typically, the surfactant has an HLB value between14 or about 14 and 20 or about 20, for example, 14, 15, 16, 17, 18, 19,20, about 14, about 15, about 16, about 17, about 18, about 19 or about20. Exemplary of suitable surfactants are sugar ester surfactants, suchas sucrose fatty acid ester (SFAE) surfactants. Typically, thesurfactant is a natural surfactant, for example, a surfactant that isGRAS (generally recognized as safe) certified by the FDA and/or Koshercertified.

The liquid concentrates further contain a polar solvent, such as water(e.g., filtered water), or other edible aqueous liquid (e.g., propyleneglycol or glycerin), or combination thereof, typically a high amount ofthe polar solvent, for example, between 60% or about 60% and 80% orabout 80%, by weight (w/w), of the concentrate, typically between at orabout 60% and at or about 79%, by weight of the concentrate.

Typically, the concentrates further contain one or more additionalingredients. Exemplary of additional ingredients that can be included inthe concentrates are preservatives, non-polar solvents, co-surfactants,emulsion stabilizers, pH adjusters and flavoring agents.

The non-polar compounds in the concentrates and dilution compositionsare contained in micelles. These micelles, containing the non-polarcompound surrounded by the one or more surfactants, allow dispersion ofthe non-polar compound among polar solvents, for example, when theconcentrates are diluted to form aqueous liquid dilution compositions.The micelles containing the non-polar compounds typically have a smallor relatively small particle size, for example, less than 1000 nm orabout 1000 nm, less than 500 nm or about 500 nm, typically less than 300nm or about 300 nm, typically less than 200 nm or about 200 nm, forexample, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150 or 200 nm. Smallerparticle size correlates with clarity of the aqueous liquid dilutioncompositions containing the diluted concentrates. For example, a liquidwith a smaller particle size is more clear than a liquid with a largerparticle size. Small particle size also can contribute to otherdesirable properties, for example, stability.

A number of factors, including ingredients, their relativeconcentrations, and methods for making the concentrates, affect theparticle size of the compositions, and other desirable properties of thecompositions, such as clarity. In particular, the nature of thesurfactant, particularly the HLB of the surfactant, and the relativeconcentrations of polar solvent (e.g., water), surfactant and thenon-polar compound, contribute to small particle size, and the clarityof the aqueous liquid dilution compositions. Typically, several of theseparameters and properties are related to one another. For example,several of the parameters contribute to the particle size, typicallysmall particle size, of the compositions. Particle size contributesdirectly to clarity of the aqueous liquid dilution compositionscontaining the concentrates. Particle size also can relate to otherproperties, for example, stability, lack of “ringing” and/or precipitateformation of the aqueous liquid dilution compositions containing theconcentrates.

Accordingly, properties of the ingredients and their relativeconcentrations in the concentrates are important for the ability of theconcentrate to yield desirable dilution compositions. Provided aremethods for formulating the liquid nanoemulsion concentrates.Determining the appropriate ingredients, and relative concentrationsthereof, that will yield dilution compositions having desirableproperties, is performed using provided methods for formulating theliquid concentrates.

a. Formulating the Liquid Concentrates

In the provided formulation methods, the concentrates are formulated byselecting ingredients and concentration ratios of the ingredients thatyield compositions having one or more desired properties. Whenformulating the concentrates, selected ingredients and startingconcentrations are used to make initial concentrates, which areevaluated and modified, if necessary.

As a first step in formulating the provided concentrates, one or moreinitial concentrates are made and evaluated for desired properties. Forthis step, ingredients are selected, for example, from among theingredients described herein. The ingredients generally includesurfactants, polar solvents, non-polar active ingredients, and otheringredients. A starting concentration (weight percentage) of eachselected ingredient is selected from within the appropriateconcentration range for that ingredient or category of ingredient, forexample, the appropriate concentration range for the surfactant. In somecases, the initial concentrate is formulated based on the ingredients,and concentrations thereof, of an existing concentrate, having one ormore desired properties.

The initial concentrate(s) then is/are made, using the methods formaking the concentrates, provided below, adding each ingredient at itsstarting concentration at the appropriate step. In one example, morethan one initial concentrate, e.g., multiple initial concentrates, eachhaving a different concentration of one or more ingredients, is made,and compared. In one example, multiple initial concentrates are producedto test various representative concentrations within an appropriateconcentration range for one or more particular ingredient.

In a typical example, the initial concentrate is made by including atleast one surfactant, such as from among the surfactants describedherein, that has an HLB value between 14 or about 14 and 20 or about 20,at a starting concentration within the concentration range of between16% or about 16% and 30% or about 30%, by weight (w/w), of theconcentrate; at least one non-polar compound, at a startingconcentration within the concentration range of between 5% or about 5%and 10% or about 10%; and a polar solvent, at a starting concentrationof between 60% or about 60% and 80% or about 80%, and typically betweenat or about 60% and at or about 79%, by weight. In one example, theinitial concentrate further includes other ingredients, for example,preservative(s), co-surfactant(s), and/or other ingredients as describedherein.

After making the initial concentrate(s), the concentrate(s) is evaluatedfor one or more desired properties, for example, the ability to formdilution compositions (e.g., clear dilution compositions or dilutioncompositions having a particular turbidity value, particle size or otherproperty). The ability to form dilution compositions having one or moreproperties is determined by diluting the concentrate in aqueous medium,for example, diluting the concentrate in the aqueous medium at adilution factor of between 1:10 or about 1:10 and at most 1:1000 orabout 1:1000, typically between 1:10 or about 1:10 and 1:500 or about1:500, for example, at a dilution between 1:10 or about 1:10 and up to1:250 or about 1:250, for example, diluted between 1:10 or about 1:10,1:20 or about 1:20, 1:25 or about 1:25, 1:50 or about 1:50, 1:100 orabout 1:100, 1:200 or about 1:200, 1:250 or about 1:250, or up to 1:500or about 1:500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50,1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100, 1:110,1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180, 1:190, 1:200, 1:210,1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280, 1:290, 1:300, 1:350,1:400, 1:500, or according to other dilutions provided herein, andassessing the clarity, turbidity value, particle size, color, smell,taste, safety, stability, “ringing” or forming of precipitates, presenceof crystals or other desired property of the resulting dilutioncomposition.

After evaluation, the ingredients, and/or concentrations thereof, can beadjusted in order to generate the desired properties in the finalconcentrate. Typically, the concentration of the non-polar compound, thesurfactant, and/or the polar solvent is the concentration that isadjusted after evaluating the initial concentrate. Similarly, whenformulating multiple initial concentrates, one or more of the non-polarcompound, surfactant and polar solvent concentration is/are varied amongthe multiple initial concentrates. In some cases, following evaluation,it can be determined that additional ingredients (not included in theinitial formulation) are needed or desirable for achieving the desiredproperties of a particular concentrate. This process can be repeateduntil a concentrate having the desired property or properties isgenerated.

i. Ingredients and Typical Concentration Ranges

Each of the provided concentrates contains a non-polar compound, suchas, but not limited to, the exemplary non-polar compounds describedherein below. Typically, the non-polar compound is a non-polar activeingredient, for example, an oil-based active ingredient such as apolyunsaturated fatty acid (PUFA), a coenzyme Q or a phytochemical. Forformulating the initial concentrate, the starting concentration of thenon-polar compound typically is a concentration chosen from within aconcentration range of between 5% or about 5% and 10% or about 10% (w/w)of the concentrate, such as a starting concentration of 5% or about 5%,6% or about 6%, 7% or about 7%, 8% or about 8%, 9% or about 9%, or 10%or about 10% (w/w) of the concentrate. The non-polar compound typicallyis added as part of an oil phase, according to the provided methods formaking the concentrate.

The initial concentrate further contains at least one surfactant, whichcan be added to the water phase or the oil phase, and typically has anHLB value of between 14 or about 14 and 20 or about 20, for example, 14,15, 16, 17, 18, 19, or 20, or about 14, about 15, about 16, about 17,about 18, about 19, about 20, typically between at or about 15 and at orabout 18, including, but not limited to sugar ester surfactants (whichtypically are sucrose fatty acid surfactants containing monoesters) andanalogs and derivatives thereof, typically a natural surfactant, whichis safe and/or approved for human consumption. The surfactants include aSFAE or mixtures thereof or the SFAE(s) and a PEG-derivative of VitaminE or an analog thereof.

Typically, the starting concentration of the surfactant is chosen fromwithin a concentration range of between 16% or about 16% and 30% orabout 30% (w/w), for example, 16% or about 16%, 17% or about 17%, 18% orabout 18%, 19% or about 19%, 20% or about 20%, 21% or about 21%, 22% orabout 22%, 23% or about 23%, 24% or about 24%, 25% or about 25%, 26% orabout 26%, 27% or about 27%, 28% or about 28%, 29% or about 29%, or 30%or about 30%, by weight (w/w), of the concentrate, such as, for example,17.75%, 20.25%, 20.5%, 22.7%, or 25.2% (w/w) of the concentrate. Whenthe concentrate contains a mixture of sucrose fatty acid esters and aPEG-derivative of Vitamin E as surfactants, the compositions typicallycontain at least 1% or at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%or 10% sucrose fatty esters.

In one example, the concentration range of the surfactant is between 16%or about 16% and 26% or about 26%, by weight (w/w), of the concentrate,such as, for example, between 17% or about 17% and 25% or about 25%(w/w) of the concentrate; between 18% or about 18% and 26% or about 26%(w/w) of the concentrate; between 16% or about 16% and 18% or about 18%(w/w) of the concentrate; such as, for example, 18% or about 18% (w/w)of the concentrate, 20% or about 20% (w/w) of the concentrate, 23% orabout 23% (w/w) of the concentrate, or 25% or about 25% (w/w) of theconcentrate. In another example, the concentration range of thesurfactant is between 17% or about 17% and 26% or about 26% (w/w) of theconcentrate. In another example, the concentration range of thesurfactant is between 18% or about 18% and 25% or about 25% (w/w) of theconcentrate. In another example, the concentration range of thesurfactant is between 18% or about 18% and 20% or about 20% (w/w) of theconcentrate. In another example, the concentration range of thesurfactant is between 17% or about 17% and 20% or about 20% (w/w) of theconcentrate. In another example, the concentration range of thesurfactant is between 16% or about 16% and 20% or about 20% (w/w) of theconcentrate.

The concentrates further contain polar solvents (e.g., water, or otheredible polar solvent, e.g., propylene glycol and glycerin), typically ahigh concentration of the polar solvent, which is added to the waterphase. Typically, the starting concentration of polar solvent is chosenfrom within a concentration range of between 60% or about 60% and 80% orabout 80% (w/w) of the concentrate, for example, 60% or about 60%, 61%or about 61%, 62% or about 62%, 63% or about 63%, 64% or about 64%, 65%or about 65%, 66% or about 66%, 67% or about 67%, 68% or about 68%, 69%or about 69%, 70% or about 70%, 71% or about 71%, 72% or about 72%, 73%or about 73%, 74% or about 74%, 75% or about 75%, 76% or about 76%, 77%or about 77%, 78% or about 78%, 79% or about 79%, 80% or about 80% (w/w)of the concentrate, such as, for example, 68.29%, 68.7865%, 69.02%,74.25%, 71.49%, 71.74%, or 75.8165% (w/w) of the concentrate. In oneexample, the concentration range of the polar solvent is between 65% orabout 65% and 80% or about 80% (w/w) of the concentrate. In anotherexample, the concentration range of the polar solvent is between 65% orabout 65% and 75% or about 75% (w/w) of the concentrate or between 65%or about 65% and 76% or about 76% (w/w) of the concentrate.

One or more, typically more than one, additional ingredients can beadded to the initial concentrate. For example, the concentratestypically contain at least one preservative, typically a naturalpreservative, for example, benzyl alcohol. Exemplary of other additionalingredients that can be added to the concentrates, including the initialconcentrates, are emulsion stabilizers, for example, a blend of gums; anon-polar solvent for the non-polar compound, for example, an oil otherthan the non-polar compound, for example, vitamin E oil or flax seedoil; a pH adjuster, for example, citric acid or phosphoric acid; one ormore flavoring agents, for example, D-limonene or lemon oil; aco-surfactant, for example, a phospholipid (e.g., phosphatidylcholine).

The appropriate concentration ranges for the additional ingredients aredescribed in individual sections below. Typically, the concentration ofthe additional ingredients depends, in part, on the concentrations ofthe non-polar active ingredient, the surfactant and the polar solvent.Typically, the concentrations of these three ingredients (surfactant,polar solvent and non-polar compound) are the focus of the formulatingmethods. For example, when it is determined that modifications toingredient concentrations in the initial concentrate should be made, ittypically is the concentrations of one or more of these threeingredients that are adjusted.

In one example, it can be desirable to add one or more of the additionalingredients after evaluation of the initial concentrate, for example, inorder to improve the concentrate with respect to one or more desiredproperties.

ii. Evaluation of the Initial Concentrate

The formulation methods further include analysis of the initialconcentrate based on one or more desired properties, for example,properties of an aqueous liquid dilution composition containing thediluted concentrate, for example, clarity, color, smell, taste, safety,stability, “ringing” or forming of precipitates and/or the presence ofcrystals. For example, the methods typically include analyzing theability of the initial concentrate to form a clear liquid upon dilutionin an aqueous medium, such as by analysis of the clarity/turbidity ofthe resulting aqueous liquid dilution composition containing the initialconcentrate.

For evaluation of properties of the aqueous liquid dilution composition,the initial concentrate is diluted into an aqueous medium, typicallywater or another polar solvent, for example, at a dilution factor ofbetween 1:10 or about 1:10 and at most 1:1000 or about 1:1000, typicallybetween 1:10 or about 1:10 and 1:500 or about 1:500, for example, at adilution between 1:10 or about 1:10 and up to 1:250 or about 1:250, forexample, diluted between 1:10 or about 1:10, 1:20 or about 1:20, 1:25 orabout 1:25, 1:50 or about 1:50, 1:100 or about 1:100, 1:200 or about1:200, 1:250 or about 1:250, or up to 1:500 or about 1:500, for example,1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75,1:80, 1:85, 1:90, 1:95, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:160,1:170, 1:180, 1:190, 1:200, 1:210, 1:220, 1:230, 1:240, 1:250, 1:260,1:270, 1:280, 1:290, 1:300, 1:350, 1:400, 1:500, or any other dilution,such as others provided herein. Typically, clarity of the aqueous liquiddilution composition containing the diluted initial concentrate isevaluated using one or more approaches. Additionally, other propertiescan be evaluated, for example, smell and/or taste properties of theliquid, for example, when the non-polar compound is a polyunsaturatedfatty acid (PUFA), particularly fish oil or algae oil, whether theaqueous liquid dilution composition smells “fishy” can be evaluatedempirically.

(1) Clarity

In one example, dilution of the provided concentrates in aqueous mediayields clear liquids. The clarity of the aqueous liquid dilutioncomposition containing the initial concentrate can be evaluated by oneor more of a plurality of approaches, such as by empirical observation,by measuring particle size and/or by measuring the turbidity value ofthe liquid.

In one example, the concentrates can be diluted to form clear liquids(or liquids that are equal in clarity to known liquids), by addingbetween 0.05 grams (g) or about 0.05 g and 10 g or about 10 g of theconcentrate, typically between 0.05 g and 5 g, for example, 0.05 g, 0.06g, 0.07 g, 0.08 g, 0.09 g, 0.1 g, 0.2 g, 0.3 g, 0.4 g, 0.5 g, 0.6 g, 0.7g, 0.8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, or 10 g ofthe concentrate, to 8 fluid ounces, about 8 fluid ounces, or at least 8fluid ounces or at least about 8 fluid ounces, for example 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 ormore fluid ounces, of aqueous medium, for example, water, forming aclear aqueous liquid dilution composition that contains the non-polarcompound. In another example, the concentrates can be diluted to formclear aqueous liquid dilution compositions by adding between 1 mL orabout 1 mL and 10 mL or about 10 mL of the concentrate, for example, 1mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL or 10 mL of theconcentrate to 8 fluid ounces, about 8 fluid ounces, or at least 8 fluidounces or at least about 8 fluid ounces, for example 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or morefluid ounces, of aqueous medium, for example, water, forming a clearaqueous liquid dilution composition that contains the non-polarcompound.

In another example, the concentrate can be diluted in aqueous medium toform a clear aqueous liquid dilution composition when at least 25 mg orabout 25 mg, typically at least 35 mg, for example, 35, 40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170,180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 325,350, 375, 400, 425, 450, 475, 500, 550, 600, 700, 800, 900, 1000, 1500,2000 mg, or more, of the non-polar active ingredient, is contained in atleast 8 fluid ounces or at least about 8 fluid ounces of aqueous liquiddilution composition, for example, a beverage, for example, water.

In another example, the concentrate can be diluted in an aqueous mediumto form a clear aqueous liquid dilution composition at a dilution factorof between 1:10 or about 1:10 and at most 1:1000 or about 1:1000,typically between 1:10 or about 1:10 and 1:500 or about 1:500, forexample, at a dilution between 1:10 or about 1:10 and up to 1:250 orabout 1:250, for example, diluted between 1:10 or about 1:10, 1:20 orabout 1:20, 1:25 or about 1:25, 1:50 or about 1:50, 1:100 or about1:100, 1:200 or about 1:200, 1:250 or about 1:250, or up to 1:500 orabout 1:500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50,1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100, 1:110,1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180, 1:190, 1:200, 1:210,1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280, 1:290, 1:300, 1:350,1:400 or at most 1:500. In another example, the clear liquid is formedat dilutions less dilute than 1:10 of the concentrate.

The provided liquid nanoemulsion concentrates can be formulated usingany non-polar compound for dilution in an aqueous medium. In oneexample, the concentrates can be diluted in aqueous medium, for example,over a wide dilution range to form clear liquids, for example, at adilution factor of between 1:10 or about 1:10 and at most 1:500 or about1:500, typically between 1:10 or about 1:10 and at most 1:1000 or about1:1000, typically between 1:10 or about 1:10 and 1:500 or about 1:500,for example, at a dilution between 1:10 or about 1:10 and up to 1:250 orabout 1:250, for example, diluted between 1:10 or about 1:10, 1:20 orabout 1:20, 1:25 or about 1:25, 1:50 or about 1:50, 1:100 or about1:100, 1:200 or about 1:200, 1:250 or about 1:250, or up to 1:500 orabout 1:500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50,1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100, 1:110,1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180, 1:190, 1:200, 1:210,1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280, 1:290, 1:300, 1:350,1:400 or at most 1:500.

Clarity of the aqueous liquid dilution composition can be evaluatedusing one or more of a plurality of approaches, for example,quantitatively, for example, by measuring particle size and/or bymeasuring the turbidity value of the liquid, or qualitatively, byempirical evaluation. In one example, a particular quantitative orqualitative clarity value is desired. In another example, it is desiredthat the aqueous liquid dilution composition is as clear as, less clearor more clear than another liquid, for example, an aqueous liquiddilution composition made according to the provided methods or abeverage, for example, a beverage or other aqueous medium that does notcontain the concentrate. For example, an aqueous liquid dilutioncomposition, containing the liquid concentrate diluted in a beverage,can be as clear or about as clear as the same beverage, containing noconcentrate. The evaluation can be done qualitatively, for example byempirical observation, or qualitatively, for example, by calculatingparticle size and/or turbidity value (NTU) for the liquid(s).

(2) Empirical Evaluation

The relative clarity/turbidity of the aqueous liquid dilutioncomposition containing the diluted concentrate (e.g., initialconcentrate) can be assessed qualitatively by observation. In oneexample, a liquid is considered clear if it does not have a cloudyappearance and/or if no particles are visible when looking at the liquidwith the naked eye. Clarity can be assessed empirically by comparison toother liquids, for example, water, fruit juice, soda and/or milk. Forexample, it can be desirable that the liquid is as clear or about asclear as water or another liquid, for example a beverage. For example,it can be desirable that the liquid (containing the liquid concentratediluted in an aqueous medium, for example, a beverage) is as clear orabout as clear as the aqueous medium not containing the liquidconcentrate. In a related example, it can be desired that there is nosubstantial difference, for example, no observable difference, betweenthe aqueous liquid dilution composition containing the concentrate andthe aqueous medium without the concentrate. A clear liquid is notnecessarily colorless, for example, a yellow liquid that contains novisible particles or cloudiness can be considered clear.

(3) Particle Size

Alternatively, the clarity of the aqueous liquid dilution compositioncontaining the diluted concentrate (e.g., initial concentrate) can beassessed by measuring the particle size of the liquid. Methods formeasuring particle size are known and any method for measuring particlesize that can measure particle sizes in the appropriate ranges asdescribed below, can be used.

Particle size can be analyzed by commercial services, for example, fromDelta Analytical Instruments, Inc, such as using a light-scatteringanalyzer, for example, a dynamic light scattering analyzer, for example,the Horiba® LB-550, which can measure particle sizes within a range of0.001 micron to 6 micron and uses a Fourier-Transform/IterativeDeconvolution technique for reporting data and can measure sampleconcentrations from ppm to 40% solids; the Horiba® LA-920, which is alaser light-scattering instrument having an He—Ne laser and a tungstenlamp and can determine particle sizes from 0.02 micron to 2000 micronusing Mie Theory; or other analyzers available from Delta AnalyticalInstruments, Inc.

Alternatively, the particle size can be measured microscopically, forexample, by viewing the liquid under a microscope, for example, at 640×magnification. With this method, particle size can be quantified bycomparing to a measuring device, for example, a ruler, which is visiblewhen viewing the liquid under the microscope. If any particles areobservable at this magnification, they are measured by comparison to themeasuring device. At a magnification of 640×, for example, any particlethat is about 25 nm, 25 nm, or greater than 25 nm are visible, whileparticle sizes smaller than 25 nm typically are not visible.

Typically, it is desired that the aqueous liquid dilution compositionshave a particle size less than 200 nm or less than about 200 nm, forexample, 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140,150, 160, 170, 180, 190, or 200 nm. Typically, it is desired that theaqueous liquid dilution compositions have a particle size less than 100nm or about 100 nm, less than 50 nm or about 50 nm, or less than 25 nmor about 25 nm. Typically, the particle size of the aqueous liquiddilution composition containing the concentrate is between 5 nm or about5 nm and 200 nm or about 200 nm, typically between 5 nm or about 5 nmand 50 nm or about 50 nm.

(4) Turbidity Measurement

Alternatively, clarity of the liquid can be analyzed by taking anoptical turbidity measurements, which indicates the level of cloudinessor haziness of a liquid, which correlates to size/number of particles insuspension in the liquid. The more clear a particular liquid, the lowerits turbidity value.

Turbidity can measured optically, for example, by using a nephelometer,an instrument with a light and a detector. The nephelometer measuresturbidity by detecting scattered light resulting from exposure of theliquid to an incident light. The amount of scattered light correlates tothe amount of particulate matter in the liquid. For example, a beam oflight will pass through a sample with low turbidity with littledisturbance. Other methods for measuring turbidity are well known andcan be used with the provided methods and compositions.

The units of a turbidity value measured with a nephelometer areNephelometric Turbidity Units (NTU). In one example, it is desired thatthe aqueous liquid dilution composition containing the dilutedconcentrate has low turbidity, for example, a turbidity value (NTU) of30 or about 30; or an NTU value of less than 30 or about 30, forexample, less than 29 or about 29, less than 28 or about 28, less than27 or about 27, less than 26 or about 26, less than 25 or about 25, lessthan 24 or about 24, less than 23 or about 23, less than 22 or about 22,less than 21 or about 21, less than 20 or about 20, less than 19 orabout 19, less than 18 or about 18, less than 17 or about 17, less than16 or about 16, less than 15 or about 15, less than 14 or about 14, lessthan 13 or about 13, less than 12 or about 12, less than 11 or about 11,less than 10 or about 10, less than 9 or about 9, less than 8 or about8, less than 7 or about 7, less than 6 or about 6, less than 5 or about5, less than 4 or about 4, less than 3 or about 3, less than 2 or about2, less than 1 or about 1; or 29 or about 29, 28 or about 28, 27 orabout 27, 26 or about 26, 25 or about 25, 24 or about 24, 23 or about23, 22 or about 22, 21 or about 21, 20 or about 20, 19 or about 19, 18or about 18, 17 or about 17, 16 or about 16, 15 or about 15, 14 or about14, 13 or about 13, 12 or about 12, 11 or about 11, 10 or about 10, 9 orabout 9, 8 or about 8, 7 or about 7, 6 or about 6, 5 or about 5, 4 orabout 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0 or about 0. Inanother example, the turbidity value of the aqueous liquid dilutioncomposition is less than 200 or less than about 200, for example, 200,175, 150, 100, 50, 25 or less.

In another example, it is desirable that the aqueous liquid dilutioncomposition contains a turbidity value that is comparable, for example,about the same as, the same as, or less than or greater than, theturbidity value of another liquid, for example, a beverage notcontaining the liquid concentrate or an aqueous liquid dilutioncomposition made by the provided methods.

iii. Selecting a Formulation and Modifying Formulations

After evaluation of the initial concentrate(s), either a particularformula is chosen or one or more modifications is made to the initialconcentrate formula based on the results of the evaluation. When aninitial concentrate does not display one or more desired properties,e.g., to the desired extent, based on the evaluation, the concentrationof one or more ingredients can be adjusted and another initialconcentrate made, in order to repeat the process until a concentratewith the desired properties is made. For modification of the initialconcentrate, the amount of the polar solvent, surfactant and/ornon-polar active ingredient can be adjusted, e.g., to anotherconcentration within the appropriate concentration range. Alternativeingredients also can be chosen. In one example, modification of theinitial concentrate involves the addition of one or more additionalingredients. For example, if evaluation reveals that the oil and waterphases of the concentrate or aqueous liquid dilution compositioncontaining the diluted concentrate are separating, an emulsionstabilizer can be added to the formulation. In another example, aco-surfactant can be added to help emulsify the components of theconcentrate. In another example, the phase (oil phase or water phase),to which a particular ingredient is added, is modified. For example, theformulation can be modified to change whether the surfactant is added tothe oil phase or the water phase.

In one example, when evaluation of the initial concentrate reveals thatit has desired properties, no modifications are made. In this example,the formula of the initial concentrate is used for making theconcentrate. When two or more initial concentrates are made, forexample, with increasing concentrations of an ingredient, the formula ofone of the initial concentrates can be chosen. Which formula is chosencan be based on which formula has the most desirable property.Alternatively, desirable properties can be balanced with relativeamounts of ingredients. In one example, it is desirable to choose theformulation that uses the lowest or the highest concentration of aparticular ingredient but still provides a concentrate that yields aclear liquid upon dilution in an aqueous medium. In one example, thedesired formulation is the formulation that has the lowest concentrationof the surfactant, while still providing a concentrate that yields aclear liquid upon dilution in an aqueous medium. In another example, thedesired formulation is the formulation that has the highestconcentration of the non-polar active ingredient, while still providinga concentrate that yields a clear liquid upon dilution into an aqueousmedium. In another example, the formulation that yields the clearestliquid is desired.

In another example, however, modifications are made to the formula evenif the initial concentrate bears desired properties. For example, upondetermining that a particular concentrate formulation results in desiredproperties, it can be desirable to modify the concentration of one ormore ingredients to determine whether the same desired properties can beachieved if a higher or lower concentration of the ingredient(s) isused. For example, it can be desirable to determine the lowestconcentration of surfactant that can be used, while still generating aconcentrate with a desired property, for example, the ability to form aclear liquid upon dilution in an aqueous medium. In another example, itcan be desirable to determine the highest concentration of the non-polaringredient that can be incorporated into a concentrate, while stillmaintaining the desired property, for example, the ability of theconcentrate to form a clear liquid upon dilution in an aqueous medium.In another example, one or more additional ingredients can be addedafter making an initial concentrate with desirable properties, forexample, flavoring agents and/or pH adjusting agents.

The following sections describe ingredients used in the provided liquidnanoemulsion concentrates.

b. Non-Polar Compounds

The concentrates contain one or more non-polar compounds. Non-polarcompounds include any lipophilic or lipid soluble compounds, forexample, active ingredients, that have greater solubility in organicsolvents (e.g., ethanol, methanol, ethyl ether, acetone, and benzene)and in fats and oils, than in aqueous liquid dilution compositions, forexample, water. Typically, the non-polar compounds are poorly watersoluble, for example, water insoluble or compounds having low watersolubility. The non-polar compounds include, but are not limited to,drugs, hormones, vitamins, nutrients and other lipophilic compounds.Exemplary non-polar compounds are listed herein below. The providedmethods and compositions can be used to dilute (e.g., dissolve/disperse)any non-polar compound in aqueous medium. In one example, the non-polarcompound differs from the surfactant, for example, is not a sucrosefatty acid ester. In another example, the non-polar compound is notVitamin E. Exemplary of non-polar compounds that can be used in theprovided concentrates are:

Non-polar ingredients containing essential fatty acids, for example,polyunsaturated fatty acids (PUFAs), for example, gamma-linolenic acid(GLA), for example, borage oil and evening primrose (Oenothera biennis)oil, blackcurrant seed oil, hemp seed oil, and spirulina extract;compounds containing omega-3 fatty acids, for example, natural andsynthetic omega-3 fatty acids, for example, compounds containing omega-3polyunsaturated long-chain fatty acids, including Eicosapentaenoic acid(EPA) (20:5ω3); Docosahexaenoic acid (DHA) (22:6ω3); Eicosatetraenoicacid (24:4ω3); Docosapentaenoic acid (DPA, Clupanodonic acid) (22:5ω3);16:3 ω3; 24:5 ω3 and/or nisinic acid (24:6ω3), for example, fish oil,algae oil, hill oil, canola oil, flaxseed oil, soybean oil and walnutoil; compounds containing short-chain omega-3 fatty acids, for example,Alpha-Linolenic acid (α-Linolenic acid; ALA) (18:3ω3) and Stearidonicacid (18:4ω3), esters of an omega-3 fatty acid and glycerol, forexample, monoglycerides, diglycerides and triglycerides, esters ofomega-3 fatty acid and a primary alcohol, for example, fatty acid methylesters and fatty acid esters, precursors of omega-3 fatty acid oils, forexample, EPA precursor, DHA precursor, derivatives such aspolyglycolized derivatives or polyoxyethylene derivatives, oilscontaining the omega-3 fatty acids, for example, fish oil (marine oil),for example, highly purified fish oil concentrates, perilla oil, krilloil, and algae oil, for example, microalgae oil; compounds containingomega 6 fatty acids, for example, compounds containing Linoleic acid(18:2ω6) (a short-chain fatty acid); Gamma-linolenic acid (GLA)(18:3ω6); Dihomo gamma linolenic acid (DGLA) (20:3ω6); Eicosadienoicacid (20:2ω6); Arachidonic acid (AA) (20:4ω6); Docosadienoic acid(22:2ω6); Adrenic acid (22:4ω6); and/or Docosapentaenoic acid (22:5ω6),for example, borage oil, corn oil, cottonseed oil, grapeseed oil, peanutoil, primrose oil, for example, evening primrose Oenothera biennis) oil,blackcurrant seed oil, hemp seed oil, spirulina extract, safflower oil,sesame oil and soybean oil;

Other fatty acids, for example, triglycerides, including medium chaintriglycerides, polar lipids, for example, ether lipids, phosphoric acid,choline, fatty acids, glycerol, glycolipids, triglycerides, andphospholipids (e.g., phosphatidylcholine (lecithin),phosphatidylethanolamine, and phosphatidylinositol); saw palmettoextract; and ethyl linoleate; and herb oils, for example, garlic oilsand scordinin; short-chain saturated fatty acids (4:0-10:0), Lauric acid(12:0), Myristic acid (14:0), Pentadecanoic acid (15:0), Palmitic acid(16:0), Palmitoleic acid (16:1 ω7), Heptadecanoic acid (17:0), Stearicacid (18:0), Oleic acid (18:1 ω9), Arachidic acid (20:0);

Micronutrients, for example, vitamins, minerals, co-factors, forexample, Coenzyme Q10 (CoQ10, also called ubiquinone), ubiquinol,tumeric extract (cucuminoids), saw palmetto lipid extract (saw palmettooil) echinacea extract, hawthorn berry extract, ginseng extract, lipoicacid (thioctic acid), ascorbyl palmitate, kava extract, St. John's Wort(hypericum, Klamath weed, goat weed), extract of quercitin,dihydroepiandrosterone, indol-3-carbinol;

Carotenoids, including hydrocarbons and oxygenated, alcoholicderivatives of hydrocarbons, for example, beta carotene, mixedcarotenoids complex, lutein, lycopene, Zeaxanthin, Cryptoxanthin, forexample, beta-crytoxanthin, beta carotene, mixed carotenoids complex,astaxanthin, bixin, canthaxanthin, capsanthin, capsorubin,apo-carotenal, beta-12′-apo-carotenal, “Carotene” (mixture of alpha andbeta-carotene), gamma carotene, violerythrin, zeaxanthin, esters ofhydroxyl- or carboxyl-containing members thereof;

Fat-soluble vitamins, for example, Vitamins A, D, E and K, andcorresponding provitamins and vitamin derivatives such as esters with anaction resembling that of vitamin A, D, E or K for example; retinol(vitamin A) and pharmaceutically acceptable derivatives thereof, forexample, palmitate ester of retinol and other esters of retinol, andcalciferol (vitamin D) and its pharmaceutically acceptable derivativesthereof and precursors of vitamin D, particularly vitamin D3, d-alphatocopherol (vitamin E) and derivatives thereof, including pharmaceuticalderivatives thereof, for example, Tocotrienols, d-alpha tocopherolacetate and other esters of d-alpha tocopherol, and ascorbyl palmitate,a fat-soluble version of vitamin C;

Phytochemicals, including phytoestrogens, for example, genistein anddaidzein, for example, isoflavones, for example, soy isoflavones,flavonoids, phytoalexins, for example, Resveratrol(3,5,4′-trihydroxystilbene), red clover extract, and phytosterols;

Lipid-soluble drugs, including natural and synthetic forms ofimmunosuppressive drugs, such as Cyclosporin, protease inhibitors suchas Ritonavir, macrolide antibiotics and oil soluble anesthetics such asPropofol, natural and synthetic forms of steroidal hormones, forexample, estrogens, estradiols, progesterone, testosterone, cortisone,phytoestrogens, dehydroepiandrosterone (DHEA), growth hormones and otherhormones; and

Oil-soluble acids and alcohols, for example, tartaric acid, lactylicacid butylated hydroxyanisole, butylated hydroxytoluene, lignin,sterols, polyphenolic compounds, oryzanol, cholesterol, phytosterols,flavonoids, such as quercetin and reservatol, and diallyl disulfides.

i. Polyunsaturated Fatty Acid (PUFA)-Containing Active Ingredients

Exemplary of the non-polar compounds contained in the concentrates arecompounds containing fatty acids, for example, active ingredientscontaining polyunsaturated fatty acids (PUFAs). Fatty acids arestraight-chain hydrocarbon molecules with a carboxyl (COOH) group at oneend of the chain. PUFAs are fatty acids that contain more than onecarbon-carbon double bond in the carbon chain of the fatty acid. PUFAs,particularly essential fatty acids, are useful as dietary supplements.

Different nomenclatures can be used to describe fatty acid molecules.Lipid nomenclature, for example, 18:3 ω-3, indicates the carbon chainlength, number of double bonds and the position along the carbon chainof the first carbon-carbon double bond in a fatty acid. Using thisnomenclature, each carbon along the chain is labeled according to itsposition relative to one end of the chain. For example, the first carbonaway from the carboxylate end is named α, the second is named β, and soforth. The last carbon in the molecule (furthest from the carboxy group)always is labeled ω (or omega, or n). The number of carbons and thenumber of double bonds are listed first in the lipid name of a fattyacid, separated by a colon. For example, the name “18:3” indicates thatthe molecule has eighteen (18) carbons and three (3) double bonds.Following these numbers, the position at which the first double bondappears, relative to the last (ω) carbon, is listed. For example, thenomenclature, 18:3 ω-3 (or 18:3 omega-3; or 18:3 n-3), describes a fattyacid with eighteen (18) carbons and three (3) double bonds, the first ofwhich occurs at the third carbon away from the omega carbon.

Alternatively, chemical nomenclature can be used. The chemical name of afatty acid describes the position of each double bond. In the chemicalnaming, the carbons are numbered, beginning with 1, starting with thecarbon that is part of the carboxy (COOH) group. Thus, with thisnumbering system, the α carbon is labeled “2.” The chemical name of thefatty acid lists the first carbon (from the COOH end) to participate ineach double bond.

Certain PUFAs are called essential fatty acids because mammals,including humans, cannot synthesize them using any known chemicalpathway, and must obtain them from diet or by supplementation. (U.S.Pat. No. 6,870,077; Covington, American Family Physician (2004), 70(1):133-140). The essential PUFAs are the omega-3 (ω3; n-3) fatty acids andthe omega-6 (ω-6; n-6) fatty acids. Omega-3 and omega-6 fatty acids aremethylene interrupted polyenes, which have two or more cis double bonds,separated by a single methylene group. Exemplary of Omega-3 fatty acidsare Alpha-Linolenic acid (α-Linolenic acid; ALA) (18:3ω3) (a short-chainfatty acid); Stearidonic acid (18:4ω3) (a short-chain fatty acid);Eicosapentaenoic acid (EPA) (20:5ω3); Docosahexaenoic acid (DHA)(22:6ω3); Eicosatetraenoic acid (24:4ω3); Docosapentaenoic acid (DPA,Clupanodonic acid) (22:5ω3); 16:3 ω3; 24:5 ω3 and nisinic acid (24:6ω3).Longer chain Omega-3 fatty acids can be synthesized from ALA (theshort-chain omega-3 fatty acid). Exemplary of Omega-6 fatty acids areLinoleic acid (18:2ω6) (a short-chain fatty acid); Gamma-linolenic acid(GLA) (18:3ω6); Dihomo gamma linolenic acid (DGLA) (20:3ω6);Eicosadienoic acid (20:2ω6); Arachidonic acid (AA) (20:4ω6);Docosadienoic acid (22:2ω6); Adrenic acid (22:4ω6); and Docosapentaenoicacid (22:5ω6).

While the longer chain Omega-3 and Omega-6 essential fatty acids can besynthesized from ALA (the short-chain omega-3 fatty acid) and Linolenicacid (LA), respectively, evidence suggests that conversion of theseshort chain fatty acids in humans is slow. Thus, a major source of longchain essential PUFAs is dietary (see e.g., Ross et al. (2007) Lipids inHealth and Disease 6:21; Lands (1992) FASEB 6(8): 2530). Dietarysupplements containing PUFAs, particularly essential PUFAs, aredesirable for protection against cardiovascular disease, inflammationand mental illnesses (see e.g., Ross et al. (2007) Lipids in Health andDisease 6:21; Lands (1992) FASEB 6(8): 2530; U.S. Pat. No. 6,870,077).Evidence suggests that essential fatty acids, particularly EPA and DHA,in the form of food and nutritional supplements, play a role inpreventing a number of disease states, including cardiovasculardiseases, inflammation, mental health and behavioral diseases anddisorders (see e.g., Ross et al. (2007) Lipids in Health and Disease6:21; Lands (1992) FASEB 6(8): 2530; U.S. Pat. No. 6,870,077; Covington(2004) American Family Physician 70(1): 133-140).

Omega-9 fatty acids are non-essential PUFAs. Exemplary of omega-9 fattyacids are Oleic acid (which is monounsaturated) (18:1 ω9); Eicosenoicacid (20:1 ω9); Mead acid (20:3 ω9); Erucic acid (22:1 ω9); and Nervonicacid (24:1 ω9).

Conjugated fatty acids are PUFAs with two or more conjugated doublebonds. Conjugated fatty acids can be used as nutritional supplements.Exemplary of conjugated fatty acids are Conjugated Linoleic acid (CLA),for example, 18:2 ω7, 18:2 ω6; Conjugated Linolenic acid, for example,18:3ω6, 18:3ω5; and other conjugated fatty acids, for example, 18:3 ω3,18:4 ω3, and 20:5 ω6.

(1) Omega-3 Fatty Acid Compounds

Exemplary of the PUFA-containing active ingredients that can be used inthe provided compositions are compounds that contain one or more omega-3(ω3; n-3) fatty acids, for example, compounds containing DHA and/or EPAfatty acids, for example, marine oils for example, fish oil, krill oiland algae oil; and compounds containing ALA fatty acids, for example,flax seed oil.

Typically, oils and aqueous compositions containing long-chainedpolyunsaturated fatty acids (PUFA) are susceptible to oxidation, makingthem unstable and giving them an unpleasant taste. The ingredients andrelative concentrations thereof, as well as the methods for making theconcentrates, contribute to desirable properties of DHA/EPA-containingconcentrates. In one example, ingredients and methods minimize the“fishy” odor and/or taste of DHA/EPA compositions and increase theirstability over time. In one aspect, the compounds in the concentrateshave low oxidation, contributing to these desirable properties.

(a) DHA/EPA

Exemplary of non-polar active ingredients that contain one or moreomega-3 fatty acids, which can be used in the provided compositions, arecompounds containing DHA and/or EPA, for example, marine oil, forexample, fish oil, krill oil and algae oil. Any oil containing DHAand/or EPA can be used. In one example, the non-polar active ingredientcontains between 20% or about 20% and 40% or about 40% DHA. In anotherexample, the non-polar active ingredient contains between 25% or about25% and 35% or about 35% DHA. In another example, the non-polar activeingredient contains at least 70% or about 70%, by weight (w/w), DHA, forexample, at least 75% or about 75%, at least 80% or about 80%, at least85% or about 85%, or at least 90% or about 90%, by weight (w/w), DHA. Inanother example, the non-polar active ingredient contains between 5% orabout 5% and 15% or about 15% EPA, for example, 5, 6, 7, 8, 9, 10, 11,12, 13, 14 or 15%, by weight (w/w), EPA. In another example, thenon-polar active ingredient comprises not more than 10% or about 10% EPAor less than 10% or about 10%, EPA. In another example, the non-polaractive ingredient contains DHA and EPA, for example, DHA representing atleast 20% or about 20%, by weight of the non-polar active ingredient andEPA representing not more than 13% or about 13% of the non-polar activeingredient, for example, not more than 10% or about 10%, by weight ofthe non-polar active ingredient. In another example, the non-polaractive ingredient contains DHA, representing at least 35% or about 35%of the non-polar active ingredient and EPA representing not more than13% or about 13% of the non-polar active ingredient, for example, notmore than 10% or about 10% of the non-polar active ingredient. Inanother example, the non-polar active ingredient contains DHA and EPA,for example, DHA representing at least 70% or about 70% of the non-polaractive ingredient and EPA representing not more than 13% or about 13% ofthe non-polar active ingredient, for example, not more than 10% or about10% of the non-polar active ingredient.

(i) Fish Oils

Exemplary of the PUFA-containing non-polar active ingredients that canbe used in the provided compositions are oils derived from fish, whichcontain DHA, EPA or DHA and EPA. Particularly, cold water marine fishare a known source of Omega-3 fatty acids (U.S. Pat. No. 4,670,285).Suitable fish oil containing DHA, EPA or DHA and EPA can be obtainedfrom any of a number of commercial sources, for example, fish oilsavailable from Jedwards International, Inc., any of which can be usedwith the provided compositions.

Fish oils typically are extracted from fish tissue, for example, frozenfish tissue. In one example, the fish oil is a tasteless fish oil, forexample, a cod liver oil, which has been isolated from fish, forexample, from cod liver, and then refined and deodorized, or in someother way treated so its taste becomes neutral, for example, asdescribed in International Publication Nos. WO 00/23545 and WO2004/098311. In one example, these fish oils are isolated from frozenfish tissue by a process that minimizes oxidation. Exemplary of such atasteless fish oil is Denomega™ 100, Borregaard Ingredients, Sarpsborg,Norway; distributed by Denomega Nutritional Oils AS, Boulder, Colo.Typically, the tasteless fish oil, for example, cod liver oil, containsbetween 25% or about 25% and 35% or about 35% Omega-3 fatty acids, forexample, 34% Omega-3 fatty acids. In one example, the fish oil, forexample, the Denomega™ 100 oil, contains 13% or about 13% DHA and 13% orabout 13% EPA.

Also exemplary of the fish oils that can be included in the providedcompositions are fish oils containing high amounts of Omega-3 fattyacids, for example, high amounts of DHA. One example of such a fish oilcontains at least about 85% DHA, typically greater than 85 DHA and atleast about 90% Omega-3 fatty acids, typically greater than, 90% Omega-3fatty acids. In another example, the fish oil can contain 98% PUFA, 89%Omega-3 fatty acids, about 70% DHA, about 10% EPA, 8.9% Omega-6 fattyacids and 0.7% Omega-9 fatty acids.

Exemplary of a fish oil containing high amounts of omega-3 fatty acidsthat can used as the non-polar compound in the provided compositions isan Omega-3 Fish Oil EE (O3C Nutraceuticals, supplied by JedwardsInternational Inc., Quincy, Mass.), which contains 89% Omega-3 fattyacids, 8.9% Omega-6 fatty acids, 0.7% Omega-9 fatty acids, 0.1%saturated fatty acids, 1.0% monounsaturated fatty acids, 74.5%Docosahexanoic (DHA) fatty acids, 9.3% Eicosapentaenoic (EPA) fattyacids and 98% polyunsaturated fatty acids (PUFA). This fish oil alsocontains 0.1% (16:0) palmitic acid, 0.1% (16:1 ω7) palmitoleic acid,0.1% (18:0) stearic acid, 0.6% (18:1 ω9) oleic acid, 0.1% (18:1 ω7)oleic acid, 0.3% (18:2 ω6) linoleic acid, 0.2% (18:3 ω3) linolenic acid,0.2% (18:4 ω3) octadecatetraenoic acid, 0.1% (20:1 ω9) eicosanoic acid,0.1% (20:2 ω6) eicosadienoic acid, 0.2% (20:3 ω6) Eicosatrienoic Acid,2.4% (20:4 ω6) arachidonic acid, 0.6% (20:4 ω3) arachidonic acid, 0.1%(22:1 ω11) erucic acid, 0.6% (21:5 ω3) uncosapentaenoic acid, 0.5% (22:4ω6) docosatetraenoic acid, 5.4% (22:5 ω6) docosapentaenoic acid, 3.6%(22:5 ω3) docosapentaenoic acid and 0.9% other fatty acids.

Also exemplary of a fish oil containing high amounts of Omega-3 fattyacids that can be used in the provided compositions is Omega Concentrate85 DHA TG Ultra (O3C Nutraceuticals AS, Oslo, Norway), which containsgreater than 85% DHA (C22:6n-3) and greater than 90% total omega-3 fattyacids and is isolated from fatty fish species Eugraulidae, Clupeidae andScombridae families. This fish oil is produced by purifying andconcentrating the oils from these fish with gentle technologies toincrease the concentration of omega-3 fatty acid DHA. Any fish oilcontaining DHA and/or EPA can be used as the non-polar compound in theprovided compositions. Also exemplary of the fish oils are other fishoils made by O3C Nutraceuticals, AS and other fish oils supplied byJedwards, International, Inc.

Also exemplary of the fish oils are krill oils, made according toInternational Publication No. WO 2007/080515.

(ii) Algae Oil

Also exemplary of non-polar compounds containing Omega-3 PUFAs,particularly DHA (and optionally EPA), that can be used as the non-polarcompound in the provided compositions are oils derived frommicroorganisms, for example, oils derived from marine dinoflagellates,for example, microalgae, for example, Crypthecodinium sp, particularly,Crypthecodinium cohnii. Microalgae oils, like fish oil, are an excellentsource of omega-3 fatty acids, particularly DHA (see e.g., U.S. PatentNos. 5,397,591, 5,407,957, 5,492,938 and 5,711,983). Exemplary of oilsderived from microalgae are the oils disclosed in, and oils madeaccording to the methods described in, U.S. Patent Nos. 5,397,591,5,407,957, 5,492,938 and 5,711,983 and U.S. Publication number2007/0166411, including DHASCO® and DHASCO-S® (Martek BiosciencesCorporation).

For example, U.S. Pat. No. 5,397,591 describes, inter alia, single celledible oils (algae oils) (and methods for making the oils), whichcontain at least 70% triglycerides, which contain about 20-35% DHA andlack EPA, isolated from Crypthecodinium cohnii, preferably containingmore than 70% triglycerides, having 15-20% myristic acid; 20-25%palmitic acid; 10-15% oleic acid; 30-40% DHA and 0-10% othertriglycerides. U.S. Pat. No. 5,407,957 describes, inter alia, algae oils(and methods for making the oils) derived from Crypthecodinium cohnii,preferably containing greater than about 90% triglycerides, at least 35DHA by weight (w/w), in one example, having 15-20% myristic acid, 20-25%palmitic acid, 10-15% oleic acid, 40-45% DHA, and 0-5% other oils. U.S.Pat. No. 5,492,938 describes, inter alia, single cell edible oils (andmethods for making the oils) containing at least 70% triglycerides,which contain about 20-35% DHA and lack EPA, isolated fromCrypthecodinium cohnii, in one example, containing more than 70%triglycerides, having 15-20% myristic acid; 20-25% palmitic acid; 10-15%oleic acid; 30-40% DHA; 0-10% other triglycerides. U.S. Pat. No.5,711,983 describes, inter alia, single cell edible oils (and methodsfor making the oils) containing at least 70% triglycerides, whichcontain about 20-35% DHA and lack EPA, isolated from Crypthecodiniumcohnii, in one example, containing more than 70% triglycerides, having15-20% myristic acid; 20-25% palmitic acid; 10-15% oleic acid; 30-40%DHA and 0-10% other triglycerides.

Also exemplary of suitable microalgae oils are those disclosed, forexample, in U.S. Pat. No. 6,977,166 and U.S. Publication Number US2004/0072330. Any oil derived from dinoflagellate, for example,microalgae, which contains DHA, and optionally EPA, is suitable as analgae oil for use with the provided compositions, for example, V-Purealgae oil (Water4Life, Switzerland, which contains EPA and DHA.

(b) Flax Seed Oil—Omega 3 (ALA)

Also exemplary of the Omega-3 containing non-polar compounds used in theprovided compositions is flaxseed oil (flaxseed oil, linseed oil).Flaxseed oils, which are good sources of omega-3 fatty acids,particularly alpha-linolenic acid, have been used as nutritionalsupplements. Flaxseed oils are produced by pressing the flax seed andrefining the oil from the flax seeds. Exemplary of flaxseed oil that canbe used as the non-polar compound in the provided compositions isflaxseed oil derived from Linum usitatissimum L., for example, flaxseedoil supplied by Sanmark LLC, Greensboro, N.C. (Sanmark Limited, Dalian,Liaoning Province, China), which contains not less than (NLT) 50% C18:3alpha-linolenic acid, and further contains other fatty acids, forexample, 3-8% C16:0 Palmitic acid, 2-8% C18:0 Stearic acid, 11-24% C18:1Oleic acid, 11-24% C18:2 linoleic acid and 0-3% other fatty acids. Alsoexemplary of suitable flaxseed oil is a flaxseed oil containing 6%Palmitic acid, 2.5% stearic acid, 0.5% arachidic acid, 19% oleic acid,24.1% linoleic acid, 47.4% linolenic acid, and 0.5% other fatty acids.The fatty acid composition of flaxseed oil can vary. Any flaxseed oilcan be used as the non-polar compound in the provided compositions. Inone example, the flaxseed oil contains at least 50% alpha-linolenic acidor at least about 50% alpha-linolenic acid. In another example, theflaxseed oil contains at least 65% or about 65% or 70% or about 70%alpha-linolenic acid. Exemplary of a flaxseed containing greater than65% linolenic acid content (of total fatty acid content), for example,70-80% or 70-75%, is the flaxseed described in U.S. Pat. No. 6,870,077.

(2) Omega-6 Compounds

Also exemplary of the non-polar compounds used in the providedcompositions are compounds containing omega-6 PUFAs, for example,gamma-linolenic acid (GLA), for example, borage oil and evening primrose(Oenothera biennis) oil, blackcurrant seed oil, hemp seed oil, fungaloil and spirulina extract. Any oil containing omega-6 fatty acids can beused in the provided compositions.

(a) Borage Oil (Gamma-Linolenic Acid (GLA))

Exemplary of the omega-6 containing non-polar compounds are compoundscontaining GLA, for example, borage oil. GLA is an omega-6 PUFA, whichprimarily is derived from vegetable oils, for example, evening primrose(Oenothera biennis) oil, blackcurrant seed oil, hemp seed oil, andspirulina extract. GLA has been used as a nutritional supplement. It hasbeen proposed that GLA has a role in treating various chronic diseasesand in particular that it has anti-inflammatory effects (Fan and ChapkinThe Journal of Nutrition (1998), 1411-1414). In one example, thenon-polar active ingredient contains at least about 22% or about 22%, byweight (w/w), GLA, for example, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 50, 60%, or more, by weight (w/w),GLA.

Borage (Borago officinalis), also known as “starflower” is an herb withseeds containing high amounts of GLA. Exemplary of borage oil that isused as a non-polar active ingredient in the provided compositions isthe borage oil supplied by Sanmark LLC, Greensboro, N.C. (SanmarkLimited, Dalian, Liaoning Province, China), derived by pressing andisolating oil from the seeds of Borago officinalis L. This oil containsnot less than (NLT) 22% C18:3 gamma-linolenic acid (GLA), between 9 and12% C16:0 Palmitic acid, between 3% and 5% C18:0 Stearic acid, between15% and 20% C18:1 Oleic acid, between 35% and 42% C18:2 linoleic acid,between 3% and 5% C20:1 Ocosenoic acid, between 1% and 4% C22:1Docosenoic acid and between 0% and 4% other fatty acids. Other borageoils can be used. Other GLA-containing oils also can be used as thenon-polar compound.

(3) Saw Palmetto Extract

Also exemplary of the non-polar compounds used in the providedcompositions is saw palmetto extract, a lipophilic extract of the ripeberries of the American dwarf palm (also called Serenoa repens or Sabalserrulata), which has been used to treat genitourinary and otherdiseases and to enhance sperm production, breast size and libido, as amild diuretic, a nerve sedative, an expectorant and a digestive tracttonic, and particularly to treat benign prostate hyperplasia (BHP)(Ernst, Academia and Clinic (2002), 136; 42-53; Gordon and Shaughnessy,Complementary and Alternative Medicine (2003), 76(6); 1281-1283). Sawpalmetto extract is commercially available from a number of sources. Anysaw palmetto lipid extract can be used in the provided compositions.Exemplary of the saw palmetto extract that can be used in the providedcompositions is Saw Palmetto, Lipophilic Extract, commercially availablefrom Natural Medicinals, Inc., Felda, Fla. This Saw Palmetto LipophilicExtract is Carbon Dioxide extracted and, in one example, contains, 85.9%total fatty acids, including 0.8% Caproic acid, 2% Caprylic acid, 2.4%Capric acid, 27.1 Lauric acid, 10.3 Myristic acid, 8.1% Palmitic acid,0.2% Palmitoleic acid, 2% Stearic acid, 26.7 Oleic acid, 4.9% Linoleicacid, 0.7% linolenic acid, 0.42%; 0.42% phytosterols, including 0.42%beta Sitosterol, 0.09% Campesterol, 0.03% Stigmasterol; and 0.2%moisture. Other sources of saw palmetto extract can be used.

(4) Conjugated Linoleic Acid (CLA)

Also exemplary of the PUFA non-polar compounds that can be used in theprovided compositions are non-polar compounds containing conjugatedfatty acids. Conjugated fatty acids are PUFAs with two or moreconjugated double bonds. Conjugated fatty acids can be used asnutritional supplements. Exemplary of the active ingredients containingconjugated fatty acids are compounds containing Conjugated Linoleic acid(CLA), for example, 18:2 ω7, 18:2 ω6; Conjugated Linolenic acid, forexample, 18:3ω6, 18:3ω5; and other conjugated fatty acids, for example,18:3 ω3, 18:4 ω3, and 20:5 ω6. CLA refers to a family of linoleic acidisomers found primarily in meat and dairy products of ruminants.Typically, the CLA compounds contain a mixture of different CLA isomers,for example, C18:2 CLA c9,t11, CLA t10, c12 and other CLA isomers.Exemplary of the CLA that can be used as an active ingredient in theprovided compositions is CLA (80%) commercially available from Sanmark,LTD (Dalian, Liaoning Province, China; product code 01057-A80). This CLAis clear white to pale yellow oil and has the following fatty acidcomposition: NMT (not more than) 9.0% C16:0 Palmitic acid, NMT 4.0%Stearic acid, NMT 15.0% C18:1 Oleic acid, NMT 3.0% C18:2 Linoleic acid,NLT (not less than) 80% C18:2 CLA (including the following isomers: NLT37.5% C18:2 CLA c9,t11, 37.5% C18:2 CLA t10, c12, and NMT 5.0% other CLAisomers); and NMT 5.0% other fatty acids. Other CLA containing compoundscan be used.

ii. Coenzyme Q Active Ingredients

Exemplary of the non-polar active ingredients are compounds containingCoenzyme Q, for example, Coenzyme Q10 (also called CoQ10, ubiquinone,ubidecarenone, ubiquinol and vitamin Q10). Coenzyme Q compounds arebenzoquinone compounds containing isoprenyl units. The number ofisoprenyl units in each of the different CoQ species is indicated with anumber following CoQ. For example, CoQ10 contains 10 isoprenyl units.Coenzyme Q10 is a predominant Coenzyme Q species.

Coenzyme Q can exist in two different forms: an oxidized form and areduced form. When the oxidized form of a Coenzyme Q species is reducedby one equivalent, it becomes a ubisemiquinone, denoted QH, whichcontains a free radical on one of the oxygens in the benzene ring of thebenzoquinone. Oxidized and reduced coenzyme Q containing compounds canbe used as active ingredients in the provided compositions.

(1) Coenzyme Q10

Exemplary of the Coenzyme Q containing non-polar active ingredients thatcan be used in the provided compositions are active ingredientscontaining Coenzyme Q10. Coenzyme Q10 (also called CoQ10, ubiquinone,ubidecarenone, ubiquinol, and vitamin Q10) is a benzoquinone compoundthat contains 10 isoprenoid units. The “Q” in the name refers to Quinoneand the 10 refers to the number of isoprenoid units. CoQ10 typicallyrefers to the oxidized form of CoQ10, which also is referred to asubidecarenone, as opposed to the reduced form of CoQ10. In both, thereduced and oxidized CoQ10 are exemplary of the coenzyme Q species thatcan be used as active ingredients in the provided compositions.

CoQ10 has electron-transfer ability and is present in cellularmembranes, such as those of the endoplasmic reticulum, peroxisomes,lysosomes, vesicles and the mitochondria. A decrease in natural CoQ10synthesis has been observed in sick and elderly people. Because of thisobservation and its potent antioxidant properties, CoQ10 is used as adietary supplement and a treatment for diseases such as cancer and heartdisease. CoQ10, however, exhibits relatively poor bioavailability.

CoQ10 containing compounds are available commercially. Any CoQ10compound or reduced CoQ10 compound can be used with the providedcomposition. Exemplary of the CoQ10 compounds that can be used as activeingredients are coenzyme Q10 compounds containing greater than 98% orgreater than about 98% ubidecarenone, for example, the compound soldunder the name Kaneka Q1™ (USP Ubidecarenone) by Kaneka Nutrients, L.P.,Pasadena, Tex. The compound sold under the name Kaneka Q10™ is fermentedentirely from yeast and is identical to the body's own CoQ10 and freefrom the cis isomer found in some synthetically produced CoQ10compounds. Any CoQ10 compound can be used in the provided compositions.

iii. Phytosterol-Containing Active Ingredients

Exemplary of the non-polar compounds used as active ingredients in theprovided compositions are phytosterol (plant sterol)-containingcompounds. Plant sterols are structurally similar to cholesterol andhave been found to reduce the absorption of dietary cholesterol, whichcan affect the levels of serum cholesterol. According to the U.S. Foodand Drug Administration (FDA), two servings per day, each containing 0.4grams of plant sterols, for a total daily intake of at least 0.8 grams,as part of a diet low in saturated fat and cholesterol, may reduce therisk of heart disease. Thus, plant sterols are used in nutritionalsupplements.

Any phytosterol-containing compound can be used as an active ingredientin the provided compositions. Exemplary of the phytosterol-containingcompounds that can be used as active ingredients in the providedcompositions are compounds containing plant sterols, for example, thecompound sold under the name CardioAid™, distributed by B&D Nutritionand manufactured by ADM Natural Health and Nutrition, Decatur, Ill. Thiscompound contains Kosher, Pareve, and Halal plant sterols that areproduced under current food GMPs. The sterols are PCR negative and thematerial is derived from genetically modified organisms (GMOs). Thisphytosterol compound contains a minimum of 95% plant sterols, which caninclude up to 5 plant sterols. The compound can contain, for example,40-58% Beta sitosterol, 20-30% Campesterol, 14-22% Stigmasterol, 0-6%Brassicasterol and 0-5% Sitostanol. The compound further can containtocopherols, for example, 0-15 mg/g tocopherols. The compound is testedand is negative for microorganisms, such as Salmonella, E. coli andStaphylococcus aureus.

c. Surfactants

The provided compositions contain surfactants. For example, in additionto the non-polar compound(s), the liquid concentrates contain one ormore surfactants. In the provided methods for producing theconcentrates, the surfactant is added to the water phase, the oil phase,or to the water and the oil phase. The compositions further can containone or more co-surfactants or emulsifiers. Typically, the surfactantsare natural surfactants, for example, a surfactant that is G.R.A.S.(generally recognized as safe) by the FDA and/or Kosher certified.

The surfactants aggregate in aqueous liquids, such as in the providedcompositions (e.g., concentrates and aqueous liquid dilutioncompositions) to form micelles, which contain the non-polar compound(s).The hydrophilic portion(s) of the surfactant molecules are orientedtoward the outside of the micelle, in contact with the aqueous medium,while the hydrophobic portion(s) of the surfactant molecules areoriented toward the center of the micelle, in contact with the non-polarcompound(s), which is contained in the center of the micelle. Themicelles can contain more than one surfactant and/or co-surfactant.Properties of the provided compositions, for example, the particle sizeof the compositions and desirable properties related to the particlesize, are influenced by the choice of surfactant(s) and the relativeamount (concentration) of surfactant. For example, the HLB of thesurfactant(s) can affect particle size, clarity, taste, smell, crystalformation and other properties of the provided compositions. Similarly,the concentration of the surfactant compared with the concentration(s)of other ingredients, particularly compared with the concentration ofthe polar solvent(s) and the concentration of the non-polar compound(s),can affect various desirable properties, for example, the ability todisperse or dissolve in aqueous media, e.g., to form a clear aqueousliquid dilution composition or pleasant taste and/or smell.

Surfactants (and co-surfactants) are molecules that contain hydrophobicand hydrophilic portions. In one example, the hydrophobic portion is ahydrophobic tail and the hydrophilic portion is a hydrophilic head ofthe surfactant molecule.

The HLB value of a surfactant is derived from a semi-empirical formula;HLB values are used to index surfactants according to their relativehydrophobicity and hydrophilicity. An HLB value is a numericalrepresentation of the relative representation of hydrophilic groups andhydrophobic groups in a surfactant or mixture of surfactants. The weightpercent of these respective groups indicates properties of the molecularstructure. See, for example, Griffin, W. C. Soc. Cos. Chem. 1:311(1949).

Surfactant HLB values range from 1-45, while the range for non-ionicsurfactants typically is from 1-20. The more lipophilic a surfactant is,the lower its HLB value. Conversely, the more hydrophilic a surfactantis, the higher its HLB value. Lipophilic surfactants have greatersolubility in oil and lipophilic substances, while hydrophilicsurfactants dissolve more easily in aqueous liquids. In general,surfactants with HLB values greater than 10 or greater than about 10 arecalled “hydrophilic surfactants,” while surfactants having HLB valuesless than 10 or less than about 10 are referred to as “hydrophobicsurfactants.” HLB values are known for a number of surfactants Tables 1Aand 1B list HLB values of exemplary surfactants and co-surfactants.

Exemplary of surfactants that can be used in the provided methods andcompositions are surfactants having an HLB value of between 14 or about14 and 20 or about 20, typically between 16 or about 16 and 18 or about18, for example, 14 or about 14, 15 or about 15, 16 or about 16, 17 orabout 17, 18 or about 18, 19 or about 19, or 20 or about 20.

The surfactants typically are, and typically have an HLB value betweenat or about 14 and at or about 20. Particular examples of suitablesurfactants for use in the provided compositions include non-ionicsurfactants, such as sugar derived surfactants, including fatty acidesters of sugars and sugar derivatives. For examples, sugar fatty acidesters include fatty acid esters of sucrose, glucose, maltose and othersugars, esterified to fatty acids of varying lengths (e.g., varyingnumbers of carbons). The fatty acids typically have carbon chainsbetween 8 and 28 carbons in length, and typically between 8 and 20, orbetween 8 and 18 or between 12 and 18, such as, but not limited to,stearic acid (18 carbons), oleic acid (18 carbons), palmitic acid (16carbons), myristic acid (14 carbons) and lauric acid (12 carbons).Typically, the sugar ester surfactants are sucrose ester surfactants,typically sucrose fatty acid ester surfactants.

(1) Sucrose Fatty Acid Ester Surfactants

Sucrose fatty acid ester surfactants contain one or more sucrose fattyacid esters, which are non-ionic surfactants that contain sucrose in thehydrophilic portions and fatty acids in the hydrophobic portions. Thesucrose fatty acid esters can be made by well-known methods (see, forexample, U.S. Pat. Nos. 3,480,616, 3,644,333, 3,714,144, 4,710,567,4,898,935, 4,996,309, 4,995,911, 5,011,922 and 5,017,697 andInternational Patent Application Publication No. WO 2007/082149),typically in an esterification reaction as described below.

Because sucrose contains eight hydroxy (OH) groups, the esterificationreaction can join the sucrose molecule to one fatty acid molecule, orcan join it to a plurality of, fatty acid molecules, producing differentdegrees of esterification, e.g., mono-, di-, tri- and poly- (up toocta-) fatty acid esters, but primarily mono-, di-, and/or tri-esters.The degree of esterification can depend on conditions of esterification.The esterification reaction can be carried out with a single type offatty acid, or a plurality of fatty acids, such as fatty acids withvarying carbon chain lengths, branched and linear fatty acids, and/orsaturated or unsaturated fatty acids. The esterification reaction with asingle fatty acid can produce a single ester, and typically forms morethan one ester, such as mono- di-, tri- and/or poly-esters, formed fromone reaction. The relative amounts of mono- di- tri- and/or poly-esterscan depend on reaction conditions.

The fatty acid in the sucrose fatty acid ester can be any fatty acid,and can contain between 4 and 28 carbon atoms, typically between 8 and28 carbon atoms, and typically between 8 and 25 carbon atoms, such asbetween 8 and 18 carbon atoms, such as 8, 9, 10, 11, 12, 13, 14, 15, 16,17 and 18 carbon atoms. The fatty acid can be synthetic or naturallyoccurring, and include linear and branched fatty acids. The fatty acidsinclude, but are not limited to, myristic acid, palmitic acid, stearicacid, oleic acid, caproic acid, capric acid, myristic acid, decanoicacid and pelargonic acid.

Thus, the sucrose fatty acid ester surfactants include sucrosemonoesters, diesters, triesters and polyesters, and mixtures thereof,and typically contain sucrose monoesters. The sucrose fatty acid estersurfactants include single fatty acid esters and also includehomogeneous mixtures of sucrose esters, containing members withdifferent lengths of fatty acid carbon chain and/or members withdifferent degrees of esterification. For example, the sucrose fatty acidester surfactants include mixtures of monoesters, diesters, triesters,and/or polyesters. The sugar ester surfactants further include sucrosefatty acid ester analogs and homologs and mixtures thereof.

Sucrose fatty acid esters are compounds having the following formulashown in Scheme I below.

where each of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ independently is:

a hydroxyl (—OH) group, or

where:

each R is an alkyl group having 3-27 carbon atoms; and

when more than one of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ is

each R can be a different alkyl group (e.g., having different number ofcarbon atoms and/or different saturation), or can be the same alkylgroup.

Typically, in the provided sucrose fatty acid ester surfactants, each Rhas between 7 and 27 carbon atoms, and typically between 7 and 19 atoms,such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 carbon atomsor between 7 and 17 carbon atoms:

An alkyl group can be a straight chain or branched alkyl group, can besubstituted or unsubstituted, and can be a saturated “saturated alkylgroup,” meaning that it does not contain any alkene or alkyne groups; oran “unsaturated alkyl group,” meaning that it contains at least onealkene or alkyne group. An alkyl group that includes at least onecarbon-carbon double bond (C═C) also is referred to by the term“alkenyl,” and alkenyl groups optionally can be substituted. An alkylgroup that includes at least one carbon-carbon triple bond (C≡C) also isreferred to by the term “alkynyl,” and alkynyl groups optionally can besubstituted.

Typically, the sucrose fatty acid ester surfactants contain sucrosefatty acid monoesters, having the structure set forth in Scheme V, whereone of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ (typically X¹) is

and the other seven of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸ are each,independently, —OH. An exemplary monoester has the following structure:

where R is an alkyl group having 3-27 carbons, and typically 7-27carbons.

The sucrose fatty acid esters include blends of sucrose fatty acidesters, which typically include monoesters, and can also includediesters, triesters and polyesters, which have structures according toScheme V, above, where two (diesters), three (triesters) or more(polyesters) of X¹, X², X³, X⁴, X⁵, X⁶, X⁷ and X⁸, (and typically X¹ andX⁸) independently, are

In general, sucrose fatty acid esters, including mixtures of sucrosefatty acid esters, can have varying HLB values, such as HLB valuesranging from at or about 1 to at or about 20. The HLB value of thesucrose fatty acid ester generally depends on the degree ofesterification (e.g., the average degree of esterification in a mixtureof different esters). Typically, the lower the degree of esterification(e.g., average degree), the higher the HLB value of the sucrose fattyacid ester or mixture thereof. Exemplary sucrose esters include sucrosedistearate (HLB=3), sucrose distearate/monostearate (HLB 12), sucrosedipalmitate (HLB=7.4); sucrose monostearate (HLB=15), sucrosemonopalmitate (HLB>10); Sucrose monolaurate (HLB 15). Typically, thesucrose fatty acid ester surfactants in the provided compositions havean HLB value of between at or about 14 and at or about 20, such as at orabout 14, 15, 16, 17, 18, 19, or 20, and typically between at or about14 and at or about 18, such as, but not limited to, HLB values of at orabout 15, 16 and 17, such as, for example, sucrose ester surfactantsincluding sucrose monopalmitate, sucrose monolaurate and sucrosemonostearate.

The sugar ester surfactants include sucrose ester blends, for example,sucrose ester mixtures containing a specified amount (e.g., percent, byweight) of sucrose monoesters. Exemplary surfactants include sucroseester mixtures having at least at or about 50%, by weight (w/w),monoester, such as at or about or at least at or about 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%, by weight (w/w), sucrosemonoesters, and typically at least at or about 60%, by weight or atleast at or about 70%, by weight (w/w), monoesters. The surfactantsinclude mixtures of sucrose esters containing at least at or about 50%sucrose monoesters, mixtures of sucrose esters containing at least at orabout 60% sucrose monoesters, mixtures of sucrose esters containing atleast at or about 70% sucrose monoesters, mixtures of sucrose esterscontaining at least at or about 80% sucrose monoesters, and mixtures ofsucrose esters containing at least at or about 90% sucrose monoesters,for example, mixtures containing at or about 72% sucrose monoesters, ator about 61% sucrose monoesters, or at or about 90% sucrose monoesters.

The sucrose fatty acid ester surfactants include sucrose fatty acidmonoesters, such as sucrose monocaprylate, sucrose monodecanoate,sucrose monolaurate, sucrose monomyristate, sucrose monopalmitate,sucrose monostearate, sucrose monopelargonate, sucrose monoundecanoate,sucrose monotridecanoate, sucrose monopentadecanoate and sucrosemonoheptadecanoate. The sucrose fatty acid esters further includemixtures containing varying percentages of monoesters, diesters,triesters and polyesters, such as, but not limited to, a mixture havingat or about 72% monoesters, 23% diesters, 5% triesters and 0%polyesters; a mixture having at or about 61% monoesters, 30% diesters,7% triesters, and 2% polyesters; and a mixtures having at or about 52%monoesters, 36% diesters, 10% triesters and 2% polyesters.

The sucrose fatty acid ester surfactants include sucrose fatty acidesters sold under the trade name DK Ester®, produced by Dai-Ichi KogyoSeiyaku Co., Ltd of Japan (which, in some examples, can be producedaccording to the methods described in U.S. Pat. Nos. 4,898,935,4,996,309, 4,995,911, 5,011,922 and 5,017,697, and distributed throughMontello Inc., Tulsa, Okla., such as the F-160 and F-140 grade esterssold under the trade name DK Ester®, and sucrose esters sold under thetrade name SURFHOPE® SE PHARMA, by Mitsubishi-Kagaku Foods Corporation,distributed by Mitsubishi Chemical Performance Polymers, Inc. Thesesucrose fatty acid esters are mixtures of esters with different degreesof esterification. The sucrose fatty acid esters further include Ryotosugar esters, which are food-grade esters sold by Mitsubishi-KagakuFoods Corporation, distributed by Mitsubishi Chemical PerformancePolymers, Inc. Exemplary sucrose fatty acid esters sold under the tradename DK Ester®, and those sold under the trade name SURFHOPE® SE PHARMAand Ryoto sugar esters, are listed in Table 1B, below. The table liststhe average degree of esterification or the fatty acid compositionwithin the mixture, and the HLB of the sucrose fatty acid estersurfactant. Any of the surfactants in Table 1B can be used. Typically,the surfactant (e.g., a surfactant listed in table 1B), has an HLB valuebetween at or about 14 and at or about 20, typically between at or about15 and at or about 18, e.g., but not limited to, those surfactants inthe table having an HLB of 15 or 16, such as the sucrose fatty acidester surfactant sold under the name DK ESTER® F-160, produced byDai-Ichi Kogyo Seiyaku Co., Ltd of Japan, and distributed throughMontello Inc., Tulsa, Okla. Other exemplary sucrose fatty acid estersurfactants are described in Youan et al., AAPS Pharma Sci 2003; 5(2)Article 22; 1-9 and in Okamoto et al., Biol. Pharm. Bull. 28(9):1689-1694 (2005).

TABLE 1B Exemplary Sucrose Fatty Acid Ester (SFAE) SurfactantsDistribution (by Average weight) Sucrose Fatty Degree of Fatty acid ofEster Acid Ester Esterification composition H.L.B. Mono:Di:Tri:Poly DKEster ® F-160 1.23 16 72% monoester; 23% diester; 5% triester DK Ester ®F-140 1.35 13 61% monoester; 30% diester; 7% triester; 2% polyester DKEster ® F-110 1.48 11 52% monoester; 36% diester; 10% triester; 2%polyester DK Ester ® F-90 1.53 9.5 45% monoester; 39% diester; 12%triester; 4% polyester DK Ester ® F-70 1.60 8 39% monoester; 45%diester; 12% triester; 4% polyester DK Ester ® F-50 1.69 6 34%monoester; 46% diester; 17% triester; 3% polyester DK Ester ® F- 3.11 211% monoester; 21% 20W diester; 14% triester; 54% polyester DK Ester ®F-10 4.85 1 0% monoester; 5% diester; 11% triester; 84% polyesterSURFHOPE ® C12 (100%) 5 32% monoester; SE PHARMA 68%di-/tri-/poly-esters J-1205 SURFHOPE ® C12 (100%) 16 81% monoester; SEPHARMA 19% di-/tri-/poly-esters J-1216 SURFHOPE ® C16 (80%); C18 16 79%monoester; SE PHARMA (20%) 21% di-/tri-/poly-esters J-1616 SURFHOPE ®C16 (70%); C18 5 30% monoester; SE PHARMA (30%) 70% di-/tri-/poly-estersJ-1805 SURFHOPE ® C16 (70%); C18 7 41% monoester; SE PHARMA (30%) 59%di-/tri-/poly-esters J-1807 SURFHOPE ® C16 (70%); C18 16 75% monoester;SE PHARMA (30%) 25% di-/tri-/poly-esters J-1816 SURFHOPE ® Sucrosestearate 3 Approximately 20% SE PHARMA (approximately 70% monoester;D-1803 stearate) approximately 80% di-/ tri-/poly-esters SURFHOPE ®Sucrose stearate 3 20% monoester; SE PHARMA (70% stearate) 80%di-/tri-/poly-esters D-1803F SURFHOPE ® Sucrose stearate 5 30%monoester; SE PHARMA (70% stearate) 70% di-/tri-/poly-esters D-1805SURFHOPE ® Sucrose stearate 7 40% monoester; SE PHARMA (70% stearate)60% di-/tri-/poly-esters D-1807 SURFHOPE ® Sucrose stearate 9 50%monoester; SE PHARMA (70% stearate) 50% di-/tri-/poly-esters D-1809SURFHOPE ® Sucrose stearate 11 55% monoester; SE PHARMA (70% stearate)45% di-/tri-/poly-esters D-1811 SURFHOPE ® Sucrose stearate 11 55%monoester; SE PHARMA (70% stearate) 45% di-/tri-/poly-esters D-1811FSURFHOPE ® Sucrose stearate 15 70% monoester; SE PHARMA (70% stearate)30% di-/tri-/poly-esters D-1815 SURFHOPE ® Sucrose stearate 16 75%monoester; SE PHARMA (70% stearate) 25% di-/tri-/poly-esters D-1816SURFHOPE ® Sucrose palmitate 15 70% monoester; SE PHARMA (80% palmitate)30% di-/tri-/poly-esters D-1615 SURFHOPE ® Sucrose palmitate 16 80%monoester; SE PHARMA (80% palmitate) 20% di-/tri-/poly-esters D-1616SURFHOPE ® Sucrose laurate 16 80% monoester; SE PHARMA (95% laurate) 20%di-/tri-/poly-esters D-1216 Ryoto S-970 Sucrose stearate 9 50% monoesterRyoto S-1170 Sucrose stearate 11 55% monoester Ryoto S-1570 Sucrosestearate 15 70% monoester Ryoto S-1670 Sucrose stearate 16 75% monoesterRyoto P-1570 Sucrose palmitate 15 70% monoester Ryoto P-1670 Sucrosepalmitate 16 80% monoester Ryoto LWA- Sucrose laurate 15 70% monoester1570 Ryoto L-1695 Sucrose laurate 16 80% monoester Ryoto OWA- Sucroseoleate 15 70% monoester 1570

(2) Production of Sucrose Esters

As noted above, methods for producing sucrose esters are well known(see, for example, U.S. Pat. Nos. 3,480,616, 3,644,333, 3,714,144,4,710,567, 4,898,935, 4,996,309, 4,995,911, 5,011,922 and 5,017,697 andInternational Patent Application, Publication No. WO 2007/082149). Thesucrose fatty acid surfactants can be produced by any well known method,and typically in an esterification reaction, for example, by reactingsucrose with a methyl ester of the desired fatty acid, such as a solventprocess, where sucrose is reacted with a methyl ester of a fatty acid inthe presence of a catalyst (e.g., potassium carbonate) and an organicsolvent (e.g., dimethyl formamide (DMF) or dimethyl sulfoxide (DMSO)),followed by purification, or in an aqueous medium process, where sucroseis mixed in a molten mixture with fatty acid salt using water without anorganic solvent and then reacted with a higher fatty acid methyl esterin the presence of a catalyst, followed by purification, and such as byany of the methods described in International Patent ApplicationPublication No. WO 2007/082149, whereby a sucrose molecule (which is adisaccharide containing one six-carbon aldo-sugar glucose linked to afive-carbon keto-sugar fructose, having the formula: C₁₂H₂₂O₁₁) isjoined to one or more fatty acids.

For example, the sucrose fatty acid ester can be produced byesterification using dimethyl formamide (DMF) as a solvent, by producinga methyl ester of the fatty acid and then reacting the methyl ester withsucrose in DMF in the presence of a catalyst (e.g., potassiumcarbonate), for example, for 4-6 hours at 83-95° C., for example, using30 to 127 parts sucrose to 30 parts methyl ester of the fatty acid(e.g., methyl stearate), 2 parts potassium carbonate and 300 partssolvent; by a similar method, but using dimethyl sulfoxide (DMSO) as thesolvent, for example, as described in U.S. Pat. No. 3,480,616; or, asdescribed in U.S. Pat. No. 3,644,333, by mixing sucrose with methylfatty acid and sodium fatty acid and previously prepared sucrose ester,using potassium carbonate as a catalyst and water as a solvent; or, asdescribed in U.S. Pat. No. 3,714,144, where sodium, potassium or lithiumsoap of the fatty acid is reacted in a molten sugar solution for two totwenty minutes under vacuum at 170-190° C., and purified, for example,as described in U.S. Pat. No. 4,710,567, by adding aqueous salt solutionfollowed by three-phase separation. In one example, the sucrose fattyacid esters are prepared and purified as described in U.S. Pat. Nos.4,898,935, 4,996,309, 4,995,911, 5,011,922 and 5,017,697, by producingthe esters by chemical catalysis, such as with the solvent process,e.g., using a DMSO solvent and potassium carbonate catalyst, or aqueoussolution method, followed by extraction and purification of the sucrosefatty acid esters, e.g., by adjusting pH, precipitation, separation andneutralization and filtration.

In another example, the sucrose fatty acid esters can be produced, asdescribed in International Patent Application Publication No. WO2007/082149, by mixing and reacting sucrose and vinyl esters of thefatty acids which can produce sucrose fatty acid ester mixtures with amonoester content of at or about 90%, and/or an acid value of lessthan 1. Briefly, this process can be carried out by dissolving sucrosein a solvent (e.g., DMSO), at a reaction temperature of between at orabout 30° C. and at or about 60° C., such as between about 40° C. and60° C. (e.g., at 60° C.), and a catalyst added and the mixture stirred,such as for 30 minutes, followed by removal of undissolved catalyst bydecanting or filtration, followed by addition of vinyl fatty acid, andreaction, such as for at or about 15 minutes, with monitoring to measureamount of vinyl fatty acid ester, for example, until the amount of vinylfatty acid ester reaches no more than at or about 10%, by weight (w/w),of the starting amount. The amount of sucrose and vinyl ester can vary.In one example, the ratio of sucrose to vinyl ester is between at orabout 2:1 and at or about 8:1. In one example, the sucrose is added at aconcentration of at or about 400 nm and the vinyl ester added at aconcentration of at or about 100 nM. The catalyst can be catalyzed by abase, such as metal oxides, metal hydroxides and metal carbonates, suchas potassium hydroxide, sodium hydroxide, potassium carbonate, sodiumcarbonate and lithium carbonate, which can be added at a concentrationof between at or about 1.5 grams/L and at or about 6 g/L of reactionvolume. In one example, the vinyl ester is vinyl stearate and thecatalyst is potassium carbonate. The resulting mixture can thenpurified, such as by vacuum distillation and addition of sodium chlorideto effect emulsification and purification methods described inInternational Patent Application Publication No. WO 2007/082149.

(3) PEG-Derivatives of Vitamin E

The SFAE(s) or mixtures thereof, can be mixed with a PEG-derivative(s)of vitamin E or analogs thereof, such that the total amount is stillwithin the range of the surfactants as described herein. Among thePEG-derived surfactants are PEG-derivatives of Vitamin E, such astocopherol and tocotrienol-derived surfactants, in which the Vitamin Emoiety represents the hydrophobic region of the surfactant, and isattached, via a linker, to another moiety, such as a polyethylene glycol(PEG) moiety. Exemplary of the Vitamin-E derived surfactants include,but are not limited to, tocopherol-derived surfactants, includingpolyalkylene glycol derivatives of tocopherol, typically polyethyleneglycol (PEG) derivatives of tocopherol, such as tocopherol polyethyleneglycol diesters (TPGD), e.g., tocopherol polyethylene glycol succinate(TPGS), TPGS analogs, TPGS homologs, and TPGS derivatives. Exemplarysurfactants also include other PEG derivatives having similarproperties, for example, PEG derivatives of sterols, e.g., a cholesterolor a sitosterol (including, for example, any of the PEG derivativesdisclosed in U.S. Pat. No. 6,632,443) and PEG-derivatives of otherfat-soluble vitamins, for example, some forms of Vitamin A (e.g.,Retinol) or Vitamin D (e.g., Vitamin D1-D5).

The surfactants include polyethylene glycol (PEG)-derivatives of VitaminE, for example, a tocopherol polyethylene glycol diester (TPGD). In oneexample, the TPGD is selected from among tocopherol sebacatepolyethylene glycol, tocopherol dodecanedioate polyethylene glycol,tocopherol suberate polyethylene glycol, tocopherol azelaatepolyethylene glycol, tocopherol citraconate polyethylene glycol,tocopherol methylcitraconate polyethylene glycol, tocopherol itaconatepolyethylene glycol, tocopherol maleate polyethylene glycol, tocopherolglutarate polyethylene glycol, tocopherol glutaconate polyethyleneglycol and tocopherol phthalate polyethylene glycol. In another example,the TPGD surfactant is a tocopherol polyethylene glycol succinate(TPGS), such as a TPGS-1000 and/or a d-α TPGS. In another example, thesurfactant is a TPGS analog. In one aspect, the surfactant is a TPGShomolog, such as, for example, a TPGS homolog that differs from a TPGSparent compound by the addition or removal of one or more methyleneunit(s), e.g., —(CH₂)_(n)—.

The PEG moieties in the PEG-derived surfactants, including the PEGmoieties in the PEG-derivatives of Vitamin E, include PEG moietiesselected from among any one or more of PEG-OH, PEG-NHS, PEG-aldehyde,PEG-SH, PEG-NH₂, PEG -CO₂H, methylated PEGs (m-PEGs) and branched PEGs,and includes PEG moieties having a molecular weight of between 200 kDaor about 200 kDa to 20,000 kDa or about 20,000 kDa, typically between200 kDa or about 200 kDa and 6000 kDa or about 6000 kDa, for example,between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa,typically between 200 kDa or about 200 kDa and 2000 kDa or about 2000kDa, between 600 kDa or about 600 kDa and 1500 kDa or about 1500 kDa, orbetween 600 kDa or about 600 kDa and 1000 kDa or about 1000 kDa.

The Vitamin E-derived surfactants (e.g., tocopherol-derived or atocotrienol-derived surfactants) include polyalkylene glycol derivativesof Vitamin E, typically polyethylene glycol (PEG) derivatives of VitaminE, for example, PEG derivatives of tocopherol or tocotrienol. SuitablePEG derivatives of Vitamin E typically contain one or more tocopherolsor tocotrienols, joined (for example, by an ester, ether, amide orthioester bond) with one or more PEG moieties, via a linker, forexample, a dicarboxylic acid linker. An exemplary surfactant is shownschematically below:

where the line between the PEG and Linker; and the line between theLinker and Vitamin E each independently represent a covalent bondselected from among an ester, ether, amide or thioester.

Typically, the Vitamin E PEG derivatives are made by joining the PEGmoiety, via esterification, to a vitamin E-linker conjugate (e.g., atocopherol-linker conjugate). In one example, the tocopherol-linkerconjugate first is formed by covalently joining (by esterification) thehydroxyl moiety of tocopherol with a dicarboxylic acid to produce anester bond. In this example, the tocopherol-linker conjugate is atocopherol ester (such as tocopherol succinate). The esterificationreaction can be performed by any of a number of known methods (see, forexample, U.S. Pat. Nos. 2,680,749, 4,665,204, 3,538,119 and 6,632,443).To make the tocopherol-PEG surfactant, the resulting tocopherol esterthen is joined (via the linker) to the PEG molecule, in anotheresterification reaction. In this example, the resulting surfactant is atocopherol polyethylene glycol diester (TPGD).

Alternatively, PEG derivatives of a tocopherol-linker ortocotrienol-linker conjugate can be made by other methods. Variousmethods known in the art for producing PEG derivatives can be used tojoin a PEG molecule to tocopherol-linker or tocotrienol-linkercompounds. For example, a tocopherol-linker conjugate can be covalentlybonded to the PEG molecule via an amide, ether or thioether bond. Forexample, a tocopherol-linker conjugate that contains an amine group canbe reacted with a PEG-NHS derivative to form an amide bond between thetocopherol-linker and the PEG molecule. A tocopherol-linker conjugatethat contains an amine group can be reacted with a PEG-aldehydederivative to form an amide bond between the tocopherol-linker and thePEG molecule. In another example, a tocopherol-linker that contains ancarboxylic acid can be activated to the corresponding acid halide andreacted with a PEG-SH derivative to form a thioester bond between thetocopherol-linker and the PEG molecule.

(a) Tocopherols and Tocotrienols

The tocopherol(s) used to make the surfactant can be any natural orsynthetic Vitamin E tocopherol, including but not limited toalpha-tocopherols, beta-tocopherols, gamma-tocopherols and deltatocopherols, either in pure forms or in heterogenous mixtures of morethan one form. Exemplary tocopherols are d-α-tocopherols andd,1-tocopherols. To make the surfactant, the tocopherol typically isesterified with a linker, for example, a dicarboxylic acid, to form atocopherol ester, which then is joined to a PEG moiety.

The tocotrienol(s) used to make the surfactants can be any natural orsynthetic Vitamin E tocotrienol, including but not limited toalpha-tocotrienols, beta-tocotrienols, gamma-tocotrienols and deltatocotrienols, either in pure forms or in heterogenous mixtures of morethan one form. Mixtures of tocopherols and tocotrienols, arecontemplated for use in the provided methods and compositions. Atocotrienol can be esterified with a linker, such as a dicarboxylicacid, before joining with a PEG moiety.

(b) PEG Moieties

The PEG used in the tocopherol-PEG derivative can be any of a pluralityof known PEG moieties. Exemplary of suitable PEG moieties are PEGmoieties having varying chain lengths, and varying molecular weights,for example, PEG 1000, PEG 200, PEG 500, and PEG 20,000. The numbersfollowing individual PEG moieties indicate the molecular weight (inkilodaltons (kDa) of the PEG moieties. The PEG moiety of thetocopherol-derived surfactant typically has a molecular weight ofbetween 200 kDa or about 200 kDa and 20,000 kDa or about 20,000 kDa,typically between 200 kDa or about 200 kDa and 6000 kDa or about 6000kDa, for example, between 600 kDa or about 600 kDa and 6000 kDa or about6000 kDa, typically between 200 kDa or about 200 kDa and 2000 or about2000 kDa, between 600 or about 600 kDa and 1500 kDa or about 1500 kDa,such as but not limited to 200, 300, 400, 500, 600, 800, and 1000 kDa.Exemplary of a PEG-derivative of tocopherol ester having a PEG moietywith 1000 kDa is TPGS-1000. Also exemplary of suitable PEG moieties arePEG moieties that are modified, for example, methylated PEG (m-PEG),which is a PEG chain capped with a methyl group. Other known PEG analogsalso can be used. The PEG moieties can be selected from among anyreactive PEG, including, but not limited to, PEG-OH, PEG-NHS,PEG-aldehyde, PEG-SH, PEG -NH₂, PEG-CO₂H, and branched PEGs.

(c) Linkers

Typically, the PEG derivatives of Vitamin E are diesters or otheresters, e.g., triesters. When the PEG derivative is a diester, thelinker joining the Vitamin E to the PEG typically is a carboxylic acid,typically a dicarboxylic acid, as in, for example, tocopherolpolyethylene glycol succinate (TPGS), where the linker is a succinicacid, and the surfactant is made by an esterification reaction joining aPEG moiety and a tocopherol ester of the dicarboxylic acid. In anotherexample, the linker is another molecule, for example, an amino acid,such as glycine, alanine, 5-aminopentanoic acid or 8-aminooctanoic acid;or an amino alcohol, such as ethanolamine.

(d) Tocopherol Polyethylene Glycol and Tocotrienol Polyethylene GlycolDiesters (Dicarboxylic Acid Esters of Vitamin E Linked to PEG)

Typically, the Vitamin E PEG derivatives are vitamin E polyethyleneglycol diesters, which are Vitamin E esters of PEG, made by joining aVitamin E ester to one or more PEG moieties by esterification. Exemplaryof the Vitamin E diesters are tocopherol polyethylene glycol diesters(TPGD) and tocotrienol polyethylene glycol diesters.

When the tocopherol or tocotrienol ester linked with the PEG moiety is atocopherol ester of a dicarboxylic acid (e.g., tocopherol succinate),the linker is a dicarboxylic acid (a carboxylic acid having two carboxygroups, e.g., succinic acid). In this example, the tocopherol ortocotrienol PEG diester is formed by esterification reaction, in whichPEG is attached to a tocopherol ester of a dicarboxylic acid.

Exemplary of dicarboxylic acids that can be used as linkers in thesetocopherol and tocotrienol PEG diester surfactants are succinic acid,sebacic acid, dodecanedioic acid, suberic acid, or azelaic acid,citraconic acid, methylcitraconic acid, itaconic acid, maleic acid,glutaric acid, glutaconic acid, fumaric acids and phthalic acids.Accordingly, exemplary of the tocopherol esters that can be esterifiedto form the PEG-derivatives are tocopherol succinate, tocopherolsebacate, tocopherol dodecanodioate, tocopherol suberate, tocopherolazelaate, tocopherol citraconate, tocopherol methylcitraconate,tocopherol itaconate, tocopherol maleate, tocopherol glutarate,tocopherol glutaconate, and tocopherol phthalate, among others.

Exemplary of the vitamin E polyethylene glycol diesters made withdicarboxylic acids are compounds having the following formula shown inscheme I below (and homologs, analogs and derivatives thereof):

where R¹, R², R³ and R⁴ each independently is H or Me; each dashed lineis independently a single or double bond; n is an integer from 1-5000; mand q each independently are 0 or 1; and p is an integer from 1-20. Inone example, the surfactant is a compound where, when each of m and q is0, p is an integer between 2-20.

In one example, the surfactant has the following formula shown in SchemeII below (including homologs, analogs and derivatives thereof):

where R¹, R², R³ and R⁴ each independently is hydrogen (H) or methyl(CH₂); the bond represented by the dashed line is either a single ordouble bond, m is an integer from 1 to 20, and n is an integer from 1 to5000.

In another example, the surfactant is a TPGS analog, such as, but notlimited to, compound other than TPGS having the formula shown in SCHEMEIII:

where R¹, R², R³ and R⁴ each independently is hydrogen (H) or methyl(CH₂); the bond represented by the dashed line is either a single ordouble bond, m is an integer from 1 to 20, and n is an integer from 1 to5000.

Exemplary of tocopherol and tocotrienol PEG diesters that can be used assurfactants in the provided compositions and methods include, but arenot limited to: tocopherol polyethylene glycol succinates (TPGS;including D-α TPGS and d,1-TPGS; see for example, U.S. Pat. No.3,102,078), tocopherol polyethylene glycol sebacate (PTS; see forexample, U.S. Pat. No. 6,632,443), tocopherol polyethylene glycoldodecanodioate (PTD; see for example, U.S. Pat. No. 6,632,443),tocopherol polyethylene glycol suberate (PTSr; see for example, U.S.Pat. No. 6,632,443) and tocopherol polyethylene glycol azelaate (PTAz;see for example, U.S. Pat. No. 6,632,443), polyoxyethanyl tocotrienylsebacate (PTrienS, for example, PTrienS-600; see for example, U.S. Pat.No. 6,632,443), as well as analogs, homologs and derivatives or any ofthe tocopherol diesters.

(e) Other Vitamin E PEG Esters

In another example, the tocopherol ester joined to the PEG to form thetocopherol PEG diester is a tocopherol ester of a tricarboxylic acid,for example, Citric acid, Isocitric acid, Aconitic acid andPropane-1,2,3-tricarboxylic acid (tricarballylic acid, carballylic acid)or a carboxylic acid having three or more carboxy groups.

In another example, the PEG derivatives of tocopherol are tocopherolpolyethylene glycol triesters (TPGT), for example, esters containing atocopherol, a linker, a PEG moiety, and an additional moiety, forexample, an additional tocopherol, a second PEG moiety, or awater-soluble group, such as a quaternary amine. In one example, whenthe triester contains two PEG moieties, each PEG moiety has a smallerchain length (and lower molecular weight) than the PEG moiety in a PEGderivative of tocopherol, having similar properties, that contains onlyone PEG chain.

(f) TPGS Surfactants

Exemplary of the tocopherol polyethylene glycol diester surfactants areTPGS, and analogs, homologs and derivatives thereof. TPGS is a naturalsurfactant that is GRAS and Kosher certified and thus, desirable for usein products designated for human consumptions, for example, beverages,food and nutritional supplements. TPGS typically has an HLB value ofbetween 16 or about 16 and 18 or about 18. Exemplary of the TPGSsurfactants is TPGS-1000, which has a PEG moiety of 1000 kDa. Exemplaryof the TPGS surfactants that can be used in the provided compositions isthe food grade TPGS surfactant sold under the name Eastman Vitamin ETPGS®, food grade, by Eastman Chemical Company, Kingsport, Tenn. Thissurfactant is a water-soluble form of natural-source vitamin E, which isprepared by esterifying the carboxyl group of crystallined-alpha-tocopheryl acid succinate with polyethylene glycol 1000 (PEG1000), and contains between 260 and 300 mg/g total tocopherol. A similarcompound can be made by esterifying the carboxyl group of the d,1 formof synthetic Vitamin E with PEG 1000. It forms a clear liquid whendissolved 20% in water. This tocopheryl polyethylene glycol is awater-soluble preparation of a fat-soluble vitamin (vitamin E), forexample, as disclosed in U.S. Pat. Nos. 3,102,078, 2,680,749 and U.S.Published Application Nos. 2007/0184117 and 2007/0141203. The PEG moietyof alternative TPGS surfactants can have a molecular weight range ofbetween about 200 kDa or 200 kDa to 20,000 kDa or about 20,000 kDa, forexample, between 600 kDa or about 600 kDa and 6000 kDa or about 6000kDa, typically between 600 kDa or about 600 kDa and 1500 kDa or about1500 kDa. Also exemplary of the TPGS surfactant that can be used in theprovided compositions is the Water Soluble Natural Vitamin E (TPGS),sold by ZMC-USA, The Woodlands, Tex. Any known source of TPGS, or anyanalog, homolog or derivative thereof, can be used.

Exemplary of TPGS analogs are compounds, other than TPGS, that aresimilar to a parent TPGS compound, but differ slightly in composition,for example, by the variation, addition or removal of an atom, one ormore units (e.g., methylene unit(s)—(CH₂)_(n)) or one or more functionalgroups.

At room temperature, TPGS typically is a waxy low-melting solid. In oneexample, the TPGS is heated prior to use, for example, to at least themelting temperature, for example, between 37° C. or about 37° C. and 41°C. or about 41° C. and the desired amount is poured out. In anotherexample, the TPGS can be added as a waxy solid to a vessel and heatedwith the heating apparatus.

Also exemplary of the surfactants are TPGS analogs, which includeVitamin E derived surfactants, including PEG derivatives of Vitamin E,including vitamin E PEG diesters, such as, but not limited to,tocopherol polyethylene glycol sebacate (PTS), tocopherol polyethyleneglycol dodecanodioate (PTD), tocopherol polyethylene glycol suberate(PTSr), tocopherol polyethylene glycol azelaate (PTAz) andpolyoxyethanyl tocotrienyl sebacate (PTrienS) as well as other PEGderivatives of Vitamin E.

ii. Concentration of the Surfactant

Typically, the concentration of the surfactant(s) (i.e., the SFAE ormixtures thereof or SFAE(s) and PEG-derivatives of vitamin E) in aparticular concentrate composition is selected, as described herein, byformulating an initial concentrate with a surfactant(s) concentrationwithin a starting concentration range, followed by evaluation of theinitial concentrate and, optionally, adjusting the surfactant(s)concentration. Alternatively, the surfactant concentration can be chosenbased on the concentration of surfactant in one or more existing liquidconcentrate formulas.

Liquid nanoemulsion concentrates provided herein contain a relativelylarge amount of a polar solvent phase compared to the smaller oil phasethat contains the non-polar compound(s) and surfactant (SFAE orSFAE/Vitamin E derivative). Emulsification (miscibility) of the oilphase and water phase can depend on the viscosity of the oil phase.Emulsification can be difficult the larger the load of surfactant in theoil phase. The Vitamin E derivatives have relatively high molecularweights, and hence emulsification can be difficult. For example, VitaminE TPGS, a PEG-derivative of Vitamin E surfactant, has a molecular weightof about 1513 g/mol. It is shown herein that addition of even arelatively small amount, for example, at least 1% or at least about 1%,2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% of a sucrose fatty acid esterrenders the oil phase more readily emulsified into the aqueous phase.Thus, in the compositions herein, sucrose fatty acid esters are added inplace of the vitamin E derivative, while maintaining the same relativepercentage of surfactant. The resulting compositions are rendered morereadily emulsified. Processing and ease of emulsification of the oil,which the SFAE is encapsulating, into water are improved.

Although the SFAE improves the processing ease, a composition containingan SFAE may not be as clear as, for example, a composition containing aconcentrate made with Vitamin E TPGS. Thus, the amount of SFAE isadjusted depending upon the clarity of the composition that is desired.For example, the SFAE containing compositions can be added to juices andsports drinks, such as GATORADE sports drink that are cloudy. The amountof SFAE can be adjusted depending upon the beverage or composition intowhich the concentrate is added.

Typically, the concentration of the surfactant is between 16% or about16% and 30% or about 30% (w/w), for example, 16% or about 16%, 17% orabout 17%, 18% or about 18%, 19% or about 19%, 20% or about 20%, 21% orabout 21%, 22% or about 22%, 23% or about 23%, 24% or about 24%, 25% orabout 25%, 26% or about 26%, 27% or about 27%, 28% or about 28%, 29% orabout 29%, 30% or about 30%, by weight (w/w), of the concentrate.Exemplary of surfactant concentrations within the appropriateconcentration range are 17.75% and 25.2% (w/w) of the concentrate.Typically, the concentration of surfactant is less than or equal to 30%or about 30% (w/w) of the concentrate. When the concentrate contains amixture of sucrose fatty acid esters and a PEG-derivative of Vitamin Eas surfactants, the compositions typically contain at least 1% or atleast about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% sucrose fattyesters.

In one example, the concentration range of the surfactant is between 17%or about 17% and 25% or about 25% (w/w) of the concentrate. In anotherexample, the concentration range of the surfactant is between 18% orabout 18% and 25% or about 25% (w/w) of the concentrate. In anotherexample, the concentration range of the surfactant is between 18% orabout 18% and 20% or about 20% (w/w) of the concentrate. In anotherexample, the concentration range of the surfactant is between 17% orabout 17% and 20% or about 20% (w/w) of the concentrate. In anotherexample, the concentration range of the surfactant is between 16% orabout 16% and 20% or about 20% (w/w) of the concentrate.

iii. HLB

Exemplary of the properties of the surfactant(s) that contribute to thedesirable properties of the compositions is the HLB(hydrophilic-lipophilic balance) of the surfactant(s). Generally, HLB isa value, derived from a semi-empirical formula, which is used to indexsurfactants according to their relative hydrophobicity/hydrophilicity.An HLB value is a numerical representation of the relativerepresentation of hydrophilic groups and hydrophobic groups in asurfactant or mixture of surfactants. The weight percent of theserespective groups indicates properties of the molecular structure. See,for example, Griffin, W. C. J. Soc. Cos. Chem. 1:311 (1949).

Surfactant HLB values range from 1-45, while the range for non-ionicsurfactants typically is from 1-20. The more lipophilic a surfactant is,the lower its HLB value. Conversely, the more hydrophilic a surfactantis, the higher its HLB value. Lipophilic surfactants have greatersolubility in oil and lipophilic substances, while hydrophilicsurfactants dissolve more easily in aqueous media. In general,surfactants with HLB values greater than 10 or greater than about 10 arecalled “hydrophilic surfactants,” while surfactants having HLB valuesless than 10 or less than about 10 are referred to as “hydrophobicsurfactants.”

HLB values have been determined and are available for a plurality ofsurfactants (e.g., see U.S. Pat. No. 6,267,985). It should beappreciated that HLB values for a given surfactant or co-surfactant canvary, depending upon the empirical method used to determine the value.Thus, HLB values of surfactants and co-surfactants provide a rough guidefor formulating compositions based on relativehydrophobicity/hydrophilicity. For example, a surfactant typically isselected from among surfactants having HLB values within a particularrange of the surfactant or co-surfactant that can be used to guideformulations. Table 1A lists HLB values of exemplary surfactants andco-surfactants.

The surfactants and HLB values set forth in Table 1A are exemplary. Anyknown surfactant or co-surfactant can be used with the providedcompositions (e.g., see U.S. Pat. No. 6,267,985), provided that it hasappropriate HLB value, such as an HLB value between at or about 14 andat or about 20. The surfactant(s) used in the provided concentratetypically has an HLB value between 14 or about 14 and 20 or about 20,for example, 14, 15, 16, 17, 18, 19, 20, about 14, about 15, about 16,about 17, about 18, about 19 or about 20. Exemplary of the surfactantsinclude, but are not limited to, non-ionic surfactants, such as sugaresters, such as sucrose fatty acid esters and mixtures thereof, such assucrose fatty acid ester mixtures including monoesters, including any ofthe sucrose fatty acid esters, analogs, homologs and derivativesthereof. Other known surfactants having HLB values between 14 or about14 and 20 or about 20, typically between about 15 and 18, also can besuitable. Typically, the surfactant is a natural surfactant, forexample, a surfactant that is G.R.A.S. (generally recognized as safe) bythe FDA and/or Kosher certified.

d. Co-Surfactants (Emulsifiers)

In one example, the liquid concentrate further contains one or moreco-surfactants (emulsifiers). For example, a co-surfactant can beincluded to improve emulsification of the active ingredient and/or thestability of the composition, for example, by preventing or slowingoxidation of the non-polar compound. Exemplary of a co-surfactant usedin the provided concentrates is a phospholipid, for example,phosphatidylcholine.

i. Phospholipids

Exemplary of the co-surfactants that can be used in the providedcompositions are phospholipids. Phospholipids are amphipathic lipid-likemolecules, typically containing a hydrophobic portion at one end of themolecule and a hydrophilic portion at the other end of the molecule. Anumber of phospholipids can be used as ingredients in the providedcompositions, for example, lecithin, including phosphatidylcholine (PC),phosphatidylethanolamine (PE), distearoylphosphatidylcholine (DSPC),phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidic acid(PA), phosphatidylinositol (PI), sphingomyelin (SPM) or a combinationthereof. Typically, the phospholipid is phosphatidylcholine (PC), whichsometimes is referred to by the general name “lecithin.” Exemplary ofthe phospholipids that can be used as co-surfactants in the providedcompositions are the phospholipids sold by Lipoid, LLC, Newark, N.J.,for example, Purified Egg Lecithins, Purified Soybean Lecithins,Hydrogenated Egg and Soybean Lecithins, Egg Phospholipids, SoybeanPhospholipids, Hydrogenated Egg and Soybean Phospholipids. SyntheticPhospholipids, PEG-ylated Phospholipids and phospholipid blends sold byLipoid, LLC. Exemplary of the phosphatidylcholine that can be used as aco-surfactant in the provided compositions is the phosphatidylcholinecomposition sold by Lipoid, LLC, under the name Lipoid S100, which isderived from soy extract and contains greater than 95% or greater thanabout 95% phosphatidylcholine.

In one example, the phospholipid, for example, PC, represents less thanor equal to 1% or about 1%, by weight (w/w) of the concentrate. In oneexample, the phosphatidylcholine represents between 0.1% or about 0.1%and 1% or about 1%, for example, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4,0.45, 0.5, 0.6, 0.65, 0.66, 0.6690, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95%,per weight (w/w), of the concentrate. In one example, the phospholipidrepresents between 0.15% or about 0.15% and 0.7% or about 0.7%, byweight (w/w) of the concentrate.

e. Polar Solvents

The compositions, including the liquid nanoemulsion concentrates and theliquid dilution compositions, further include polar solvents. Polarsolvents are well known in the art. The polarity of a solvent generallyindicates which compounds are soluble in the solvent, and with whichother solvents/liquids the solvent is miscible. Generally speaking,polar compounds are more readily solubilized in water and other polarsolvents than are non-polar compounds. Polar solvents are more likely tobe miscible with water and other polar solvents and liquids.

The polarity of a solvent can be assessed by measuring a number ofdifferent parameters according to well known methods (see, e.g.,Prizbytek, “High Purity Solvent Guide,” Burdick and JacksonLaboratories, Inc., 1980), such as by determining a property of thesolvent such as the dielectric constant, the dipole moment, or thepolarity index. For example, polar solvents generally have highdielectric constants, typically dielectric constants greater than at orabout 15 (see, e.g., Lowery et al., Mechanism and Theory in OrganicChemistry, Harper Collins Publishers, 3^(rd) ed., 1987, p. 177), such asat or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80 85, 90, orgreater than 90. For example, the dielectric constant of water is at orabout 80.10. Polar solvents generally have high polarity indices,typically greater than at or about 3 (see, e.g., Snyder, “Classificationof the solvent properties of common liquids,” J. Chromatography A,92:223-230, 1974), such as at or about 3, 4, 5, 6, 7, 8 or 9 or greaterthan 9. Polar solvents generally have large dipole moments, typicallygreater than at or about 1.4 Debye, such as at or about, 1.4, 1.5, 1.6,1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 3.0, 3.5, 4 or greaterthan 4 Debye (see, e.g., “CRC Handbook of Chemistry and Physics,” Lide,ed., 82^(nd) edition, CRC Press, 2001, p. 15(14)-15(18)). Other methodsof assessing solvent polarity are known in the art, including, but notlimited to, the Kosower Z scale (Kosower, “An introduction to physicalorganic chemistry,” Wiley, 1969, p. 293), the donor number and donoracceptor scale (Gutmann, “Solvent effects on the reactivities oforganometallic compounds,” Coord. Chem. Rev., 18:225-255, 1976), and theHildebrand solubility parameters (see, e.g., Giddings et al., “Highpressure gas chromatography of nonvolatile species. Compressed gas isused to cause migration of intractable solutes,” Science, 162:67-73,1968).

Polar solvents include polar protic solvents and polar aprotic solvents.A polar protic solvent (e.g., water, methanol, ethanol) contains ahydrogen atom attached to an electronegative atom, such that thehydrogen has a proton-like character and/or the bond between thehydrogen and electronegative atom is polarized. Polar aprotic solvents,on the other hand, (e.g., acetone, acetonitrile), generally do notcontain positively polarized hydrogen atoms.

The polar solvents in the provided compositions typically are polarprotic solvents, including, but not limited to, water; alcohols,including, but not limited to, dihydric alcohols (e.g., glycols, e.g.,propylene glycol, ethylene glycol, tetraethylene glycol, triethyleneglycol, trimethylene glycol), which contain two hydroxyl groups,trihydric alcohols (e.g., glycerin, butane-1,2,3-triol,pentane-1,3,5-triol, 2-amino-2-hydroxymethyl-propane-1,3-diol), whichcontain three hydroxyl groups, monohydric alcohols (e.g., methanol,ethanol, propanol, isopropanol, n-butanol and t-butanol) and otheralcohols; and acids, including but not limited to acetic acid and formicacid. Other polar solvents include, but are not limited to, acetone,acetonitrile, butyl acetate, dimethylformamide, dimethyl sulfoxide,dioxane, ethyl acetate, tetrahydrofuran and hexamethylphosphorictriamide. Typically, the polar solvent is water, or is an alcohol thattypically contains two or more hydroxyl groups, such as a trihydric ordihydric alcohol, such as, but not limited to, glycerol and propyleneglycol. The polar solvents further include low molecular weightpolyethylene glycols (PEGs), such as PEGs having a molecular weight notmore than at or about 600 kDa, such as between at or about 200 kDa andat or about 600 kDa, typically not more than at or about 400 kDa, forexample, not more than 200 kDa.

In one example, the polar solvent has a dielectric constant greater thanat or about 15, and typically between at or about 20 and at or about 80,such as at or about 80.1, 46.53, or 28.67. In another example, the polarsolvent has a polarity index between at or about 3 and at or about 9. Inanother example, the dipole moment of the polar solvent is between 1.5and 3, and typically between at or about 1.8 and 2.8, such as 1.9, 2.6,and 2.2 (for dielectric constants of solvents, see, for example,Landolt-Bornstein, New Series IV/17, Static Dielectric Constants of PureLiquids and Binary Liquid Mixtures, Springer, 2008; and CRC Handbook ofChemistry and Physics,” Lide, ed., 82^(nd) edition, CRC Press, 2001; fordipole moment of solvents, see, for example, CRC Handbook of Chemistryand Physics,” Lide, ed., 82^(nd) edition, CRC Press, 2001) and forpolarity indices of solvents, see, for example, Snyder, “Classificationof the solvent properties of common liquids,” J. Chromatography A,92:223-230, 1974).

The amount of the polar solvent typically is a high concentration, forexample, within a concentration range of between 60% or about 60% and80% or about 80%, by weight (w/w), of the concentrate, for example, 60%or about 60%, 61% or about 61%, 62% or about 62%, 63% or about 63%, 64%or about 64%, 65% or about 65%, 66% or about 66%, 67% or about 67%, 68%or about 68%, 69% or about 69%, 70% or about 70%, 71% or about 71%, 72%or about 72%, 73% or about 73%, 74% or about 74%, 75% or about 75%, 76%or about 76%, 77% or about 77%, 78% or about 78%, 79% or about 79%, or80% or about 80% (w/w) of the concentrate. Exemplary of polar solventconcentrations in the provided liquid concentrates are 69.02%, 71.49%,71.74%, 75.8165%, 74.25%, 68.7865%, and 68.29% (w/w) of the concentrate.In one example, the concentration range of the polar solvent is between65% or about 65% and 80% or about 80% (w/w) of the concentrate. In oneexample, the concentration range of the polar solvent is between 65% orabout 65% and 75% or about 75% (w/w) of the concentrate, or between 65%or about 65% and 76% or about 76%, by weight (w/w), of the concentrate,or between 68% or about 68% and 75% or about 75%, by weight (w/w), ofthe concentrate.

In the provided methods for making the concentrates, the polar solvent(e.g., water, propylene glycol or glycerin) is added to the water phase.In one example, the polar solvent is water, e.g., purified water, suchas water that is purified prior to adding it to the concentrate formula,for example, by charcoal filter, ion exchange, reverse osmosis, UVsterilization and/or filtering using a filter, for example, a 50-100micron filter. Typically, when a filter is used, it is an end point ofuse filter, which filters the water before it reaches the tank in theprovided process. Alternatively, previously filtered water can be addedto the concentrates.

f. Preservatives and Sterilizers

In one example, the provided liquid concentrate further comprises one ormore preservatives (or preservatives) and/or sterilizers. Thepreservative(s) can be included to improve the stability of theconcentrate, and the compositions made by diluting the concentrate, overtime. Preservatives, particularly food and beverage preservatives, arewell known. Any known preservative can be used in the providedcompositions. Exemplary of the preservatives that can be used in theprovided compositions are oil soluble preservatives, for example, benzylalcohol, Benzyl Benzoate, Methyl Paraben, Propyl Paraben, antioxidants,for example, Vitamin E, Vitamin A Palmitate and Beta Carotene.Typically, a preservative is selected that is safe for humanconsumption, for example, in foods and beverages, for example, a GRAScertified and/or Kosher-certified preservative, for example, benzylalcohol.

The preservative typically represents less than 1%, less than about 1%,1% or about 1%, by weight (w/w), of the liquid nanoemulsion concentrateor between 0.1% or about 0.1% and 1% or about 1%, by weight (w/w), ofthe concentrate, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%,0.725%, 0.75%, 0.8%, 0.9%, 1%, about 0.1%, about 0.2%, about 0.3%, about0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about1%, by weight (w/w), of the liquid concentrate.

g. Emulsion Stabilizers (Co-Emulsifier)

In one example, the provided liquid concentrates further contain one ormore emulsion stabilizers (co-emulsifiers), which can be used tostabilize the liquid nanoemulsion concentrate and/or the aqueouscompositions containing the diluted concentrates. In one example, theemulsion stabilizer increases the viscosity of the liquid concentrate.In one example, one or more emulsion stabilizers is added, duringformulation, after evaluation of an initial concentrate, particularly ifthe oil and water phases of the initial concentrate (or the aqueousliquid dilution composition resulting from dilution of the initialconcentrate) appear to be separating. Addition of the emulsionstabilizer can prevent separation of the oil and water phases.

Exemplary of an emulsion stabilizer that can be used in the providedcompositions is a composition containing a blend of gums, for example,gums used as emulsifying agents, for example, a blend containing one ormore of xanthan gum, guar gum and sodium alginate, for example, theemulsion stabilizer sold under the brand name SALADIZER®, available fromTIC Gums, Inc. (Belcamp, Md.). Other gums can be included in theemulsion stabilizer, for example, gum acacia and sugar beet pectin.Other blends of similar gums can also be used as emulsion stabilizers.

The emulsion stabilizer can be added to the water phase, the oil phase,and typically to the water and the oil phase, during formation of theliquid concentrates. In one example, the emulsion stabilizer is added tothe water phase at a concentration, such that it represents less than 1%or about 1% w/w of the liquid concentrate. In one example, the emulsionstabilizer is added to the water phase for a final concentration ofbetween 0.1% or about 0.1% and 1% or about 1%, for example, 0.1%, 0.12%,0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.25%, 0.3%,0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%,0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1% w/w of the liquid concentrate. In oneexample, the emulsion stabilizer is added to the oil phase such that itrepresents less than 0.1% or about 0.1%, for example, between 0.01% orabout 0.01% and 0.1% or about 0.1%, for example, 0.01%, 0.02%, 0.03%,0.04%, 0.05%, 0.06%, 0.061%, 0.062%, 0.063%, 0.0635%, 0.07%, 0.08%,0.09% or 0.1%, by weight (w/w) of the concentrate. In one example, theemulsion stabilizer is added to the water phase and the oil phase, forexample, at a concentration within the oil and water phase concentrationranges listed above. In one such example, the emulsion stabilizerrepresents less than 1%, for example, between 0.01% or about 0.01% and1% or about 1% (w/w), emulsion stabilizer, for example, 0.01%, 0.02%,0.03%, 0.04%, 0.05%, 0.06%, 0.061%, 0.062%, 0.063%, 0.0635%, 0.07%,0.08%, 0.09%, 0.1%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%,0.19%, 0.2%, 0.25%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%,0.37%, 0.38%, 0.39%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%, by weight(w/w), of the liquid concentrate.

h. Non-Polar Solvents

In one example, the liquid concentrates further contain a non-polarsolvent, for example, an oil. Typically, the non-polar solvent isincluded in the composition in addition to the non-polar activeingredient, and is used to dissolve the non-polar active ingredient. Inone example, the solvent is an oil that is not contained in thenon-polar active ingredient. When a non-polar solvent is included in theconcentrate, it typically is used to dissolve the non-polar compoundbefore mixing with the other ingredients, for example, before mixingwith the other oil phase ingredients. In one example, use of a non-polarsolvent reduces the crystal size and/or increase the clarity of theaqueous liquid dilution composition containing the diluted concentrate.Exemplary of non-polar solvents that can be used in the providedconcentrates are oils (in addition to the non-polar active ingredient),for example, Vitamin E oil, flaxseed oil, CLA, Borage Oil, D-limonene,Canola oil, corn oil, MCT oil and oat oil. Other oils also can be used.Exemplary of the Vitamin E oil, used as a non-polar solvent in theprovided compositions, is the oil sold by ADM Natural Health andNutrition, Decatur, Ill., under the name Novatol™ 5-67 Vitamin E(D-alpha-Tocopherol; ADM product code 410217). This Vitamin E oilcontains at least 67.2% Tocopherol and approximately 32.8% soybean oil.

In one example, the concentration of the non-polar solvent is within aconcentration range of between 1% or about 1% and 10% or about 10%, forexample, 1%, 2%, 3%, 3.25%, 3.5%, 3.75%, 4%, 5%, 5.25%, 5.5% or 5.75%,w/w, of the concentrate. In another example, the concentration is withinthe concentration range of between 3% or about 3% and 6% or about 6%,w/w, of the liquid concentrate. In another example, it is between 3.75%and 5.25% w/w, of the liquid concentrate.

i. Flavors

In one example, the concentrate further contains one or more flavors orflavoring agents, for example, any compound to add flavor to theconcentrate and/or to the aqueous liquid dilution composition containingthe diluted concentrate, for example, the food or beverage containingthe concentrate. Several flavors are well known. Any flavor can be addedto the concentrates, for example, any flavor sold by Mission Flavors,Foothill Ranch, Calif. Exemplary of flavors that can be used are fruitflavors, such as guava, kiwi, peach, mango, papaya, pineapple, banana,strawberry, raspberry, blueberry, orange, grapefruit, tangerine, lemon,lime and lemon-lime; cola flavors, tea flavors, coffee flavors,chocolate flavors, dairy flavors, root beer and birch beer flavors,methyl salicylate (wintergreen oil, sweet birch oil), citrus oils andother flavors. Typically, the flavors are safe and/or desirable forhuman consumption, for example, GRAS or Kosher-certified flavors.Exemplary of flavoring agents that can be used in the compositions arelemon oil, for example lemon oil sold by Mission Flavors, FoothillRanch, Calif.; and D-limonene, for example, 99% GRAS certifiedD-Limonene, sold by Florida Chemical, Winter Haven, Fla. Typically, theflavor is added, using the provided methods, to the nanoemulsionconcentrates after combining the oil and water phases. Alternatively,flavor(s) can be added to the water and/or oil phase directly.

Typically, the concentration of flavoring agent added to the providedconcentrates is less than 5% or about 5%, typically less than 1% orabout 1%, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,0.9%, 0.37% or 0.525%.

j. pH Adjusters

In one example, one or more pH adjusters is added to the providedconcentrates, typically to the emulsion that is formed after combiningthe water and oil phases according to the provided methods. Inparticular, the pH adjuster is used in compositions containing water.Alternatively, the pH adjuster can be added, at an appropriateconcentration to achieve a desired pH, to the oil phase and/or the waterphase. Typically, the pH adjuster is added to adjust the pH of theconcentrate to within a range of 2.0 or about 2.0 to 4.0 or about 4.0.One or more of a plurality of pH adjusting agents can be used.Typically, the pH adjusting agent is safe for human consumption, forexample, GRAS certified. Exemplary of a pH adjuster is citric acid, forexample, the citric acid sold by Mitsubishi Chemical, Dublin, Ohio.

Typically, the concentration of pH adjuster added to the providedconcentrates is less than 5% or about 5%, typically less than 1% orabout 1%, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,0.9%, 0.28% or 0.19%.

2. Powder Forms of the Compositions

The compositions also can be provided in powder form, i.e., powder thatis made by converting the provided nanoemulsion concentrates into apowder, using one of several well-known methods (e.g., spray-dryingand/or milling). The powder compositions include, but are not limitedto, coated or uncoated swallowable or chewable tablets, dry powders inhard or soft gelatin capsules, and dry powders in individual or multipleuse packages for reconstituted suspensions or sprinkles. Preferablesolid dosage forms are coated or uncoated swallowable or chewabletablets. Suitable methods for manufacturing the powder compositions arewell known in the art.

Additionally, the powder composition can further comprise at least oneexcipient. Excipients include, but are not limited to, diluents(sometimes referred to as fillers) including, for example,microcrystalline cellulose, mannitol, lactose, calcium phosphate,dextrates, maltodextrin, starch, sucrose, and pregelatinized starch;disintegrants including, for example, crospovidone, sodium starchglycolate, croscarmellose sodium, starch, pregelatinized starch, andcarboxymethylcellulose sodium; binders including, for example, starch,hydroxypropyl cellulose, hydroxypropylmethyl cellulose, pregelatinizedstarch, guar gum, alginic acid, acacia, carboxymethylcellulose sodium,and polyvinyl pyrrolidone; glidants including, for example, colloidalsilicon dioxide and talc; and lubricants/antiadherents including, forexample, magnesium stearate, calcium stearate, stearic acid, sodiumstearyl fumarate, glyceryl monostearate, hydrogenated vegetable oil, andtalc. In one particular example, the excipients are selected from anyone or more of maltodextrin and gum acacia.

The powder forms can be used for any convenient dosage amount of thenon-polar compound. Generally, the level of non-polar compound can beincreased or decreased according to the judgment of the physician,pharmacist, pharmaceutical scientist, or other person of skill in theart. The amount of the remaining non-active ingredients can be adjustedas needed.

Typically, the concentration of the excipients is within a concentrationrange of between 50% or about 50% and 85% or about 85%, for example, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85 ormore, %, by weight, of the free flowing powder.

In one example, the powder form is a free-flowing powder. Free-flowingpowders can be obtained using techniques well known in the art, such as,but not limited to, spray drying, freeze drying or absorption plating.In one example, in order to achieve a free flowing powder, the proteinderivative is formulated with an excipient such as lactose or starch.For example, the formulation can be a spray-dried lactose formulation(see e.g., U.S. Pat. No. 4,916,163).

The methods for forming the powders include spray drying. Spray-dryingprocesses and spray-drying equipment are described generally in Perry'sChemical Engineers' Handbook, pages 20-54 to 20-57 (Sixth Edition 1984).More details on spray-drying processes and equipment are reviewed byMarshall, “Atomization and Spray-Drying,” 50 Chem. Eng. Prog. Monogr.Series 2 (1954), and Masters, Spray Drying Handbook (Fourth Edition1985). Methods for spray drying are well known (see, e.g. U.S. Pat. Nos.5,430,021 and 6,534,085 and U.S. Application Publication No. US2007/0184117). In general, spray drying is used to dry a heated liquidby passing it through hot gas. One or more spray nozzles is used toatomize the liquid in a cooling tower or chamber. As the material isatomized (sprayed), the surface tension causes a uniform sphericalparticle to form, which is passed through the cooling chamber andhardens into a solid intact sphere. The spray dried particles can bebetween at or about 0.5 microns and at or about 100 microns, andtypically are less than at or about 10 microns, typically less than ator about 5 microns, and typically less than at or about, or at or about,1 micron.

Provided are methods for spray drying the liquid nanoemulsioncompositions to form powder compositions. In the spray drying methods,the liquid nanoemulsion compositions can be heated, e.g. to atemperature between at or about 100 and at or about 150° F., typicallybetween 110° F. and 140° F., e.g. at or about 110, 115, 120, 125, 130,135 or 140° F. The compositions can be mixed while heating, such as withany of the mixers described herein, for example, homogenizers (e.g.reversible homogenizers and piston-driven homogenization).

For spray-drying, one or more excipients are mixed with a polar solvent,typically water, and heated, e.g. to a temperature between at or about100° F. and at or about 150° F., typically between 110° F. and 140° F.,e.g. at or about 110, 115, 120, 125, 130, 135 or 140° F. In one example,the excipient is mixed with water in an amount of one part excipient (byweight) to two parts water (by weight). The excipient-solvent (e.g.water) mixture can be mixed while heating, e.g. using any of the mixersdescribed herein, for example, homogenizers (e.g. reversiblehomogenizers and piston-driven homogenization) with heating during themixing. The heated liquid nanoemulsion composition and the heatedwater-excipient mixture then are mixed together, such as by transferringone mixture to the other, e.g. by any of the transfer means providedherein. Typically, the two mixtures are homogenized, e.g. with areversible homogenizer or piston-driven homogenizer or any otherhomogenizer. The homogenized mixture then is subject to spray dryingusing a spray dryer.

Exemplary of the spray dryers are cyclone spray dryers. During spraydrying with cyclone spray dryers, the homogenized mixture is pumped intoan atomizing device where it is broken into small droplets. Upon contactwith a stream of hot air, the moisture is removed very rapidly from thedroplets while still suspended in the drying air. The dry powder isseparated from the moist air in cyclones by centrifugal action. Thecentrifugal action is caused by the great increase in air speed when themixture of particles and air enters the cyclone system. The dense powderparticles are forced toward the cyclone walls while the lighter, moistair is directed away through the exhaust pipes. The powder settles tothe bottom of the cyclone where it is removed through a dischargingdevice. Sometimes the air-conveying ducts for the dry powder areconnected with cooling systems which admit cold air for transport of theproduct through conveying pipes. Cyclone dryers have been designed forlarge production schedules capable of drying ton-lots of powder perhour.

As will be appreciated by one of skill in the art, the inlet temperatureand the outlet temperature of the spray drier are not critical but willbe of such a level to provide the desired particle size, of less than ator about 1 micron, and to result in a powder that has a desiredproperty. Typically, the ability of the free flowing powder to yield aclear (or relatively clear) liquid dilution composition upon dilution inan aqueous medium is the desired property that is evaluated. In thisregard, the inlet and outlet temperatures are adjusted depending on themelting characteristics of the liquid nanoemulsion components and thecomposition of the homogenized liquid nanoemulsion concentrate/excipientmixture. The inlet temperature is between at or about 60° C. and at orabout 170° C. with outlet temperatures between at or about 40° C. to ator about 120° C. Preferably inlet temperatures are from at or about 90°C. to at or about 120° C. and outlet temperatures are from at or about60° C. to at or about 90° C. The flow rate which is used in the spraydrying equipment will generally be at or about 3 mL per minute to at orabout 15 mL per minute. The atomizer air flow rate will very betweenvalues of at or about 25 L per minute to at or about 50 L per minute.Commercially available spray dryers are well known to those of skill inthe art, and suitable settings for any particular dispersion can bereadily determined by one of skill in the art without undueexperimentation. Operating conditions such as inlet temperature andoutlet temperature, feed rate, atomization pressure, flow rate of thedrying air, and nozzle configuration can be adjusted in accordance withthe manufacturer's guidelines.

In some examples, the dry powder is stored into a capsule form or ispressed into a tablet. For use as tablets, the compositions typicallycontain multiple other excipients. These excipients include tabletdisintegrants, such as the corn starch, glidants, such as the silicondioxide, and lubricants such as the magnesium stearate. Ordinarily thesecompositions contain minor amounts by weight of glidants and lubricants,e.g., each two percent (2%) or less by weight. Tablet disintegrants areoptionally present, and, if present, are included in sufficient amountsto assure that the tablet disintegrates upon ingestion. Accordingmaterials, such as corn starch are employed at concentrations of fromabout zero to about 30 percent by weight of the composition.

Free flowing powders also can be used to administer the active agent byinhalation using a dry powder inhaler. Such dry powder inhalerstypically administer the active agent as a free-flowing powder that isdispersed in a patient's air-stream during inspiration. In order toachieve a free flowing powder, the active agent is typically formulatedwith a suitable excipient such as lactose or starch. For example, such adry powder formulation can be made, for example, by combining thelactose with the active agent and then dry blending the components.Alternatively, if desired, the active agent can be formulated without anexcipient. The pharmaceutical composition is then typically loaded intoa dry powder dispenser, or into inhalation cartridges or capsules foruse with a dry powder delivery device. Examples of dry powder inhalerdelivery devices include Diskhaler (GlaxoSmithKline, Research TrianglePark, N.C.) (see, e.g., U.S. Pat. No. 5,035,237); Diskus(GlaxoSmithKline) (see, e.g., U.S. Pat. No. 6,378,519; Turbuhaler(AstraZeneca, Wilmington, Del.) (see, e.g., U.S. Pat. No. 4,524,769);Rotahaler (GlaxoSmithKline) (see, e.g., U.S. Pat. No. 4,353,365) andHandihaler (Boehringer Ingelheim). Further examples of suitable DPIdevices are described in U.S. Pat. Nos. 5,415,162, 5,239,993, and5,715,810 and references cited therein.

3. Liquid Dilution Compositions Containing the Diluted Concentrates

Among the compositions provided herein are liquid dilution compositions,typically aqueous liquid dilution compositions, containing the non-polarcompounds. The aqueous liquid dilution compositions are made by dilutingthe provided liquid nanoemulsion concentrates into aqueous media, forexample, beverages, for example, water, flavored water, soda, milk,juices, including fruit juices, sauces, syrups, soups, sports drinks,nutritional beverages, energy drinks, vitamin-fortified beverages, orany beverage.

In one example, the aqueous liquid dilution compositions containsbetween 0.05 grams (g) or about 0.05 g and 10 g or about 10 g, typicallybetween 0.05 g and 5 g, of the liquid concentrate per 8 fluid ounces orabout 8 fluid ounces, at least 8 fluid ounces or at least about 8 fluidounces, or less than 8 fluid ounces or less than about 8 fluid ounces,or per serving size, of the aqueous medium, for example, 0.05 g, 0.06 g,0.07 g, 0.08 g, 0.09 g, 0.1 g, 0.2 g, 0.3 g, 0.4 g, 0.5 g, 0.6 g, 0.7 g,0.8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, or 10 g ofthe concentrate per 8 fluid ounces, about 8 fluid ounces, or at least 8fluid ounces or at least about 8 fluid ounces of the aqueous medium, forexample 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35,40, 45, 50, 100, 200 or more fluid ounces, of aqueous medium.

In another example, the aqueous liquid dilution composition containsbetween 1 mL or about 1 mL and 10 mL or about 10 mL of the liquidconcentrate, for example, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8mL, 9 mL or 10 mL of the concentrate, per 8 fluid ounces, about 8 fluidounces, at least 8 fluid ounces or at least about 8 fluid ounces, orless than 8 fluid ounces or less than about 8 fluid ounces, or perserving size, of the aqueous medium, for example 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or morefluid ounces, of aqueous medium.

In another example, the aqueous liquid dilution composition contains atleast 10 mg or about 10 mg, typically at least 25 mg or about 25 mg,typically at least 35 mg, of the non-polar compound, for example, thenon-polar active ingredient, per 8 fluid ounces or about 8 fluid ounces,at least 8 fluid ounces or at least about 8 fluid ounces of the aqueousmedium, or less than 8 ounces or less than about 8 ounces, or perserving size, of the aqueous medium; for example, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 25, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110,120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250,260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550,600, 700, 800, 900, 1000, 1500, 2000 mg, or more, of the non-polarcompound per at least 8 fluid ounces or at least about 8 fluid ounces ofaqueous medium.

In another example, the aqueous liquid dilution composition contains theconcentrate diluted at a dilution factor of between 1:10 or about 1:10and at most 1:1000 or about 1:1000, typically between 1:10 or about 1:10and 1:500 or about 1:500, for example, at a dilution between 1:10 orabout 1:10 and up to 1:250 or about 1:250, for example, diluted between1:10 or about 1:10, 1:20 or about 1:20, 1:25 or about 1:25, 1:50 orabout 1:50, 1:100 or about 1:100, 1:200 or about 1:200, 1:250 or about1:250, or up to 1:500 or about 1:500, for example, 1:10, 1:20, 1:25,1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90,1:95, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180,1:190, 1:200, 1:210, 1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280,1:290, 1:300, 1:350, 1:400 or 1:500. In another example, the aqueousliquid dilution compositions contain the liquid concentrate diluted toany amount. In another example the dilution is less than 1:10 or about1:10.

Properties of the provided liquid concentrates that are diluted into theaqueous medium contribute to various properties of the providedresulting aqueous liquid dilution compositions, for example, clarity;desirability for human consumption, for example, pleasant taste, and/orsmell, for example, lack of “fishy” taste/smell, lack of “ringing” andlack of crystal formation; stability, for example, lack of oxidation,“ringing” and/or precipitation over time; and safety for humanconsumption. As described above, the liquid concentrates are formulatedaccording to the desired properties of the aqueous liquid dilutioncompositions containing the concentrates.

a. Clarity

In one example, the aqueous liquid dilution compositions are clearaqueous liquid dilution compositions or non-turbid aqueous liquiddilution compositions, for example, as determined, as described below,empirically or by measuring turbidity and/or particle size. In anotherexample, the aqueous liquid dilution compositions are not clear, or notcompletely clear. The liquids can be more or less clear, or have thesame clarity as another liquid, for example, an aqueous liquid dilutioncomposition made according to the provided methods or a beverage, forexample, a beverage that does not contain the diluted concentrate.Properties of the liquid concentrates can affect the clarity of theliquid. A number of parameters can vary the clarity of the liquids, forexample, the relative concentration of surfactant, non-polar compoundand/or water; the type of non-polar ingredient; the concentration ofexcipient(s) in the particular non-polar compound; and the purity of thenon-polar compound, for example, whether it has been standardized to ahigh purity, or whether it is an extract or a filtered extract. Forexample, an aqueous liquid dilution composition made by diluting aconcentrate containing a non-polar active ingredient that containslecithin, for example a high amount of lecithin, can be less clear thanone made with a concentrate containing a non-polar compound that doesnot contain lecithin. In another example, a liquid concentratecontaining a non-polar compound that is a filtered extract can produce aclearer aqueous liquid dilution composition when diluted than aconcentrate containing a crude extract.

i. Clarity Determined by Empirical Evaluation

In one example, the clarity/turbidity of the aqueous liquid dilutioncomposition containing the diluted concentrate is evaluatedqualitatively, by observation. In one example, a liquid can beconsidered clear if it does not have a cloudy appearance and/or if no orfew particles are visible when viewing the liquid with the naked eye orif it is the same or substantially similar in clarity to another liquid,for example, a beverage, for example, water, fruit juice, soda or milk.In some cases, the aqueous liquid dilution composition is as clear orabout as clear as water or another liquid, for example a beverage. Forexample, the liquid (containing the liquid concentrate diluted in anaqueous medium, for example, a beverage) can be as clear or about asclear as the aqueous medium not containing the liquid concentrate. In arelated example, there is no substantial difference, for example, noobservable difference, between the aqueous liquid dilution compositioncontaining the concentrate and the aqueous medium without theconcentrate. A clear liquid is not necessarily colorless, for example, ayellow liquid that contains no visible particles or cloudiness can beconsidered clear. In another example, the liquid is clear or partiallyclear or substantially clear if no crystals are visible and/or if no“ringing” is observed on the container containing the liquid.

ii. Clarity Determined by Particle Size or Number of Particles

In another example, clarity of the aqueous liquid dilution compositionis evaluated by measuring the particle size and/or number of particlesof the liquid.

In one example, the aqueous liquid dilution compositions have a particlesize less than 200 nm or less than about 200 nm, for example, 5, 10, 15,20, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130,140, 150, 160, 170, 180, 190, or 200 nm. In another example, the aqueousliquid dilution composition has a particle size less than 100 nm orabout 100 nm, less than 50 nm or about 50 nm or less than 25 nm or about25 nm. Typically, the particle size of the aqueous liquid dilutioncomposition is between 5 nm or about 5 nm and 200 nm or about 200 nm, orbetween 5 nm or about 5 nm and 50 nm or about 50 nm.

Typically, the particle size of the provided aqueous liquid dilutioncomposition containing the liquid concentrate, which contains thenon-polar compound, is smaller than the particle size of a liquidcontaining the non-polar compound (not formulated in a liquidconcentrate).

iii. Turbidity

In another example, the clarity of the liquid is evaluated and/orexpressed using a turbidity measurement, for example, NephelometricTurbidity Units (NTU), as measured using the provided methods, describedbelow. In this example, turbidity is measured optically, to get valueindicating the cloudiness or haziness of the liquid, which correlateswith particles in suspension in the liquid. The more clear a liquid is,the lower its turbidity value.

In one example, the clear aqueous liquid dilution composition has aturbidity value (NTU) of 30 or about 30; or an NTU value of less than 30or about 30, for example, less than 29 or about 29, less than 28 orabout 28, less than 27 or about 27, less than 26 or about 26, less than25 or about 25, less than 24 or about 24, less than 23 or about 23, lessthan 22 or about 22, less than 21 or about 21, less than 20 or about 20,less than 19 or about 19, less than 18 or about 18, less than 17 orabout 17, less than 16 or about 16, less than 15 or about 15, less than14 or about 14, less than 13 or about 13, less than 12 or about 12, lessthan 11 or about 11, less than 10 or about 10, less than 9 or about 9,less than 8 or about 8, less than 7 or about 7, less than 6 or about 6,less than 5 or about 5, less than 4 or about 4, less than 3 or about 3,less than 2 or about 2, less than 1 or about 1; or 29 or about 29, 28 orabout 28, 27 or about 27, 26 or about 26, 25 or about 25, 24 or about24, 23 or about 23, 22 or about 22, 21 or about 21, 20 or about 20, 19or about 19, 18 or about 18, 17 or about 17, 16 or about 16, 15 or about15, 14 or about 14, 13 or about 13, 12 or about 12, 11 or about 11, 10or about 10, 9 or about 9, 8 or about 8, 7 or about 7, 6 or about 6, 5or about 5, 4 or about 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0or about 0.

In another example, the turbidity value of the aqueous liquid dilutioncomposition is less than 200 or less than about 200, for example, 200,175, 150, 100, 50, 25 or less.

In another example, it is desirable that the aqueous liquid dilutioncomposition contains a turbidity value that is comparable, for example,about the same as, the same as, or less than or greater than, theturbidity value of another liquid, for example, a beverage notcontaining the liquid concentrate or an aqueous liquid dilutioncomposition made by the provided methods.

b. Stability

Typically, the provided aqueous liquid dilution compositions containingthe concentrates are stable, for example, free from one or more changesover a period of time, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or12 months, 1, 2, 3, 4 or more years.

In one example, the compositions are stable because they are free fromoxidation or substantial oxidation over time. In another example, theyare stable because they remain clear over time. In another example, thestable compositions remain safe and/or desirable for human consumptionover time. In one example, stability refers to the lack of precipitatesforming in the compositions over the period of time. In a relatedexample, the compositions are stable because they do not exhibit“ringing,” formation of a whitish or opaque ring around the perimeter ofthe container holding the liquid, typically at the surface of theliquid. Ringing typically is undesirable, particularly in the case of aliquid for human consumption, for example, a beverage.

In another example, the composition is stable if it does not exhibit anyvisible phase separation over a period of time, for example, after 24hours, after one week or after one month. In one example, thecompositions are stable if they exhibit one or more of these describedcharacteristics, over time, when kept at a particular temperature. Inone example, the compositions remain stable at room temperature, forexample, 25° C. or about 25° C. In another example, the compositionsremain stable at between 19° C. and 25° C. In another example, thecompositions remain stable at refrigerated temperatures, for example, 4°C. or about 4° C., or at frozen temperature, for example, at −20° C. orabout −20° C.

Stability refers to a desirable property of the provided compositions,for example, the ability of the provided compositions to remain freefrom one or more changes over a period of time, for example, 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11 or 12 months, 1, 2, 3, 4 or more years. In oneexample, the composition is stable if it is formulated such that itremains free from oxidation or substantial oxidation over time. Inanother example, the stable compositions remain clear over time. Inanother example, the stable compositions remain safe and/or desirablefor human consumption over time. In one example, stability refers to thelack of precipitates forming in the compositions over the period oftime. In a related example, stability refers to the lack of “ringing”over the period of time. In another example, the composition is stableif it does not exhibit any visible phase separation over a period oftime, for example, after 24 hours, after one week or after one month. Inone example, the compositions are stable if they exhibit one or more ofthese described characteristics, over time, when kept at a particulartemperature.

In one example, the compositions are stable at room temperature, forexample, 25° C. or about 25° C. In another example, the compositionsremain stable at between 19° C. and 25° C. In another example, thecompositions remain stable at refrigerated temperatures, for example, 4°C. or about 4° C., or at frozen temperature, for example, at −20° C. orabout −20° C.

c. Desirable Characteristics for Human Consumption

In one example, the liquid dilution composition is desirable for humanconsumption, for example, for use in a food or beverage. Differentproperties of the liquid dilution composition can contribute to itsdesirability as a consumable product. For example, taste, smell,clarity, color, crystal formation, precipitation and “ringing,” all canrelate to desirability.

In one example, the liquid dilution composition has a pleasant tasteand/or smell, for example, due to one or more flavors added to theconcentrate and/or to the aqueous medium. In another example, the liquiddilution composition containing the concentrate is free from anunpleasant taste or smell, for example, a “fishy” taste or smell. In oneexample, the concentrate smells or tastes less unpleasant, for example,fishy, compared to another aqueous liquid dilution composition.

In another example, the aqueous liquid dilution composition is desirablebecause it does not have crystals or has fewer crystals compared withanother aqueous liquid dilution composition. In another example, theaqueous liquid dilution composition is desirable because it does notexhibit ringing.

d. Safety

Typically, the aqueous liquid dilution compositions containing theconcentrates are safe for human consumption, for example, containingonly ingredients approved by the FDA for human consumption, for exampleGRAS-certified ingredients. In one example, one or more of theingredients, for example, all the ingredients, are Kosher-certified.Safety of the compositions also relates to stability over time. Lack ofor minimum oxidation of the compositions over time can contribute to thesafety of the compositions.

e. Oral Bioavailability

In one example, the non-polar compounds, for example, the non-polaractive ingredients, contained in the aqueous liquid dilutioncompositions exhibit a high or relatively high bioavailability, forexample, a bioavailability that is higher than a liquid containing thenon-polar active ingredient alone (i.e., not formulated in the liquidconcentrate). Bioavailability relates to the ability of the body toabsorb the non-polar active ingredient into a particular space, tissuecell and/or cellular compartment. Typically, non-polar activeingredients in liquids having small particle sizes are better absorbedthan those with larger particle sizes.

C. Methods for Making Liquid Nanoemulsion Concentrates ContainingNon-polar Compounds

Also provided are methods for making the liquid nanoemulsionconcentrates. General equipment and steps of the methods are detailedbelow. In one example, the general methods for making the concentratesare performed using a bench-top manufacturing process, which is used formaking relatively smaller-sized batches of the concentrates. In anotherexample, the general methods for making the concentrates are performedusing a scaled-up manufacturing processes, which is used for makingrelatively larger batches of the concentrates. The bench-top process canbe scaled up to the scaled-up process. Any concentrate made using thebench-top method can be made using the scaled-up process, by scaling upthe method.

1. Equipment for Making the Concentrates

Various equipment, for example, vessels for mixing the oil phase, waterphase and emulsion, for example, tanks; scales; mixers, includingstandard mixers and homogenizers; heating and cooling apparatuses,including water-jacketed tanks, hot plates, water baths and chillers(coolers), including recirculating coolers; transfer apparatuses, forexample, transfer means, for example, pumps, hoses, sanitary fittings;ball valves; purifiers, for example, filters, for example, carbonfilters, ion exchange equipment, reverse osmosis equipment, end-pointfilters and end product filters; evaluation means, for example, pH andtemperature meters; and other equipment, is used in various steps of theprovided methods for making the concentrates. The choice of equipmentdepends on a plurality of factors, including batch size andmanufacturing process.

2. Scales

One or more scales typically is used to measure the ingredients beforeadding them to the appropriate vessel. Alternatively, the ingredientscan be weighed in the vessel, for example, in a tank on top of a scale.

Any of a plurality of well-known, commercially sold scales can be usedto weigh the ingredients. Choice of scale(s) can depend on a number offactors, including the mass of the final concentrate being made and theingredient being weighed. In one example, multiple scales are used toweigh the various ingredients of the concentrate. In general, relativelylarger capacity (weight) scale(s) are used in making larger batches ofconcentrate while relatively smaller capacity scale(s) are used inmaking smaller batches.

Exemplary of the scales used with the provided methods to weigh theingredients are a Toledo Scale (Model GD13x/USA), a Sartorius BasicAnalytical Scale (Model BA110S) which is a basic series analytical scalewith a 110 g capacity and a resolution of 0.1 mg; and an OHAUS Scale(Model CS2000), which is a compact portable digital scale having a 2000g capacity and a resolution of 1 g.

a. Purifiers, Including Filters

Purifiers, typically more than one purifier, for example, filters, areused in the provided methods to remove impurities in the ingredientsprior to their addition to the concentrate and/or from the finalconcentrate and/or an intermediate phase of the concentrate. Forexample, when the polar solvent is water, the water typically ispurified water. In one example, one or more purifiers, for example,carbon filters, ion exchange purifiers, reverse osmosis purifiers,and/or end point filters are used to filter water, for example, citywater, prior to its addition to the water phase, for example, to removeimpurities, for example, sediment, from the water.

Exemplary of the purifiers that can be used with the provided methodsare filters, for example, 100 micron filters and carbon filters, whichare filters that use activated carbon to remove impurities by chemicaladsorption. Carbon filtering typically is used for water purificationand are particularly effective at filtering out chlorine, sediment,volatile organic compounds and other impurities. Typically, theparticles removed by carbon filters are between about 0.5 microns andabout 50 microns. Other filters are well known and can be used with theprovided methods.

Also exemplary of the purifiers that can be used in the provided methodsare reverse osmosis purifiers, which use mechanical pressure to purifyliquids, for example, water. In one example, the pressure forces thewater through a semi-permeable membrane to remove impurities.

Also exemplary of the purifiers that can be used in the provided methodsare ion exchange purifiers, for example, an ion exchange purifier usinga resin bed, for example, a zeolite resin bed, to replace salts, e.g.,cations, for example, magnesium and calcium, with other cations, forexample, sodium and potassium cations. Such purifiers can be purchased,for example, from Aquapure Filters, Clarkston, Mich.

In another example, an end product filter (e.g., a 100 micron FSIfilter, Product Number BPEM 100-5GP). This filter is used to filter anyimpurities out of the final product (e.g., the final liquid nanoemulsioncomposition). Other filters are known and can be used with the providedmethods.

b. Vessels for Mixing the Ingredients

One or more, typically two or more, vessels, for example, tanks, forexample, water-jacketed tanks; pots; and/or beakers, for example, Pyrex®beakers, are used in the provided methods to contain the ingredient(s)of the liquid concentrates, for example, during mixing and/or heating orcooling. Typically, separate vessels (an oil phase tank and a waterphase tank) are used for mixing and heating the ingredients of the oilphase and the water phase, prior to combining the two phases to form anemulsion. In another example, an additional vessel, for example, aholding and/or packaging tank, is used for holding and/or packaging theemulsion and/or for addition/mixing of additional ingredients to theemulsion.

A number of vessels are available for mixing ingredients. Typically, thevessels are cleaned, for example, rinsed, soaped and/or sanitizedaccording to known procedures, prior to use and between uses.

In one example, typically used with the bench-top process, the vessel isa container, for example, a bench-top container, for example, flasks,beakers, for example, Pyrex® beakers, vials, measuring containers,bottles and/or other bench-top containers.

In another example, typically used with the scaled-up manufacturingprocess, the vessels are tanks, for example, water phase tanks, oilphase tanks and holding/packaging tanks. Typically, the tanks areequipped with one or more mixers, for example, a standard mixer and/orhomogenizer, which are used to mix the ingredients added to the tank. Inone example, the tank further is equipped with a heating and/or coolingdevice. For example, the tank can be a water-jacketed tank. Thetemperature of the water-jacketed tank is controlled through thewater-jacket, for example, to heat the contents, for example, whilemixing.

Exemplary of the tanks that can be used with the provided methods arewater-jacketed tanks, for example, the Overly 550 Gallon water jacketedtank (Model 10576501G), which has a 550 gallon capacity and typically isused as a water-phase tank, the Schweitzers 450 gallon tank (Model#5214-C), which has a 450 gallon capacity and typically is used as anoil phase tank and the Royal 190 gallon water jacketed tank (Model9977-5), which has a 190 gallon capacity and can be used as a water oroil phase tank when mixing smaller volumes. Other tanks are well knownand can be used with the provided methods for mixing the concentrates,for example, the phases of the concentrates.

c. Mixers

Mixers are used in the provided methods to blend, mix and/or emulsifythe liquid concentrates and/or various ingredients and/or phases of theliquid concentrates. In one example, the mixers are used to keep theingredients and/or mixture circulating to maintain temperature,viscosity and/or other parameters of the mixture. Exemplary of themixers that can be used in the provided methods are standard mixers, forexample, standard mixers, which can be used, for example, to mix theingredients in the water and/or oil phases, to maintain a homogeneousmixture while heating. Exemplary of the standard mixers is a LIGHTNIN®mixer (LIGHTNIN, Rochester, N.Y.), for example, Model Numbers XJC117 andND-2. In one example, the LIGHTNIN® mixers are fixed-mount, gear drivehigh-flow mixers, for use with closed tanks. Another example of astandard mixer is a mixer sold by IKA®, for example, overhead IKA®mixers, for example, model Nos. RW-14 Basic and RE-16S, which arelaboratory stirrers and can be used to mix ingredients, for example, togenerate the oil and water phases. In one example, the mixer(s) areattached to the vessels, for example, the tanks, for example, mounted orclamped onto the tanks, for example, the top of the tanks. In anotherexample, the mixers are placed in the vessels for mixing.

Also exemplary of the mixers used with the provided methods arehomogenizers (also called shears), which typically are used to form theemulsion by emulsifying the oil and water phases after they arecombined. The homogenizers typically provide high shear dispersion ofsolids and emulsification of immiscible liquids at high shear rates.Exemplary of the homogenizers that can be used in the provided methodsare high-shear homogenizers, for example, reverse homogenizers sold byArde Barinco, Inc., Norwood, N.J., for example, Model CJ-50, which is a3600 rpm mixer having a 6 inch rotor diameter, a tip speed of 5575ft/minute and an emersion depth of 33 inches and has six separateopenings at the bottom and top, which concentrates the liquid into sixchambers, reducing the surface volume and creating a shear effect; andModel CJ-4E, which is a 10,000 rpm mixer with fan-cooled motor,optimized for 1 to 5 gallon batch sizes, having a 1.875 inch rotordiameter, a tip speed of 4920 rpm and an immersion depth of 16 inches.Other homogenizers, for example, other reversible homogenizers sold byArde Barinco Inc., can be used with the provided methods.

In one example, the homogenizer is attached to the top of the vessel,for example, the tank, for example, by clamps or by channel locks and anelectrical hoist. In another example, the homogenizer is placed in thevessel. The Arde Barinco reversible homogenizers contain axial flowimpellers, which create two distinct mixing actions, depending ondirection. Downward “vortex flow” pulls solids from top and bottom ofthe mixture, while upward “umbrella flow” controls mixing at the highestshear and recirculation rates without splashing or incorporation of air.The reversible homogenizers typically are equipped with an adjustablebaffle plate, which can be adjusted to control the type of mixing, forexample at different times during emulsification.

A number of additional mixers are well known and can be used with theprovided methods. Exemplary of the mixers that can be used with theprovided methods are shears, inline mixers/mixing, Ribbon, Plow/PaddleBlenders Forberg Mixers, Conveyors, Bag Dumps & Compactors, V-Blenders,Blade Mixers, Double Cone Mixers, Continuous Mixers, Speedflow Mixers,Batch Mixers, Double Ribbon Blenders, Paddle and Ribbon Mixers withChoppers, Plow Blenders/Turbulent Mixers, Fluidizing Forberg-TypeMixers, Air Mixers, Active Mixers, Passive Mixers, Top Entry Mixers,Side Entry Mixers, Static Mixers, Fixed Entry Mixers, PortableMixers—direct and gear drive, Sanitary Mixers, Drum Mixers, BulkContainer (IBC) Mixers, Lab Stirrers, Variable Speed Mixers, doughmixer, vertical mixer, spiral mixer, twin arm mixer, fork mixer, doublespiral mixer, all agitators, agitator mixers, Banbury Mixers, RubberMixers, Blondheim Mixers, Churn Mixers, Conical Mixers, ContinuousMixers, Disperser Mixers, Pan Mixers, Emulsifier Mixers, Hobart Mixers,Liquifier Mixers, Littleford Mixers, Meat Mixers, Plow Mixers, MixmullerMixers, Nauta Mixers, Oakes Mixers, Planetary Mixers, Pony Mixers, PUGMixers, Ribbon Mixers, Ross Mixers, Rotary Mixers, Sigma Mixers, SingleArm Mixers, Tote Bin Mixers, Tumble Mixers, Vacuum Mixers, TurbolizerMixers, Twin Shell Mixers, V-Type Mixers, Zig-Zag Mixers side armmixers, hand-held mixers, stir rods, stir bars, magnetic mixers andoverhead mixers, for example, mechanical and/or electric overheadmixers.

d. Heating Apparatuses

One or more, typically more than one, heating apparatuses are used inthe provided methods to control the temperature of the ingredients,phases and/or concentrate, typically while mixing.

In one example, the heating apparatuses are water-jackets. In thisexample, the vessels used to mix the ingredients and/or emulsify thephases are water jacketed tanks. The water jacket can be controlled, forexample, using a control panel, to adjust the temperature of thecontents of the vessel.

Alternatively, other heating apparatuses can be used to heat theingredients, phases, and/or concentrates. Exemplary of heatingapparatuses that can be used with the provided methods are immersibleand/or submersible heaters, for example, 12 KW or 13 KW sanitaryheaters, which are food-grade heaters that are immersed into the tankswhile mixing, typically for applications requiring high heat, forexample, temperatures greater than 60° C. or about 60° C., or greaterthan 80° C. or about 80° C. Also exemplary of heating apparatuses arestoves, for example, propane stoves. Also exemplary of the heatingapparatuses are hot plates, for example, the Thermolyne hot plate, modelnumber 846925 and model number SP46615. Typically, the heater is capableof heating the mixture to between 45° C. or about 45° C. and 85° C. orabout 85° C., for example, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85° C. Typically, theheater is capable of heating the mixture to 60° C. or about 60° C., forexample, providing low heat.

e. Cooling Apparatuses

One or more cooling apparatuses can be used with the provided methods,for example, to cool the ingredients during mixing, for example, tochill the mixture while emulsifying the oil and water phases. Exemplaryof the cooling apparatuses are chillers, for example, recirculatingcoolers, which can be attached to the vessel, for example, remotely orby a tank mounted in the cooler, to recirculate fluid from the tank,through the chiller and back to the tank, in order to rapidly cool andmaintain the temperature of the mixture during mixing. Exemplary of anopen-loop chiller that can be attached to the tank and used with theprovided methods are chillers sold by Turmoil, West Swanzey, N.H., forexample, open or closed-loop coolers, for example, model No. OC-1000 RO.Other cooling apparatuses are well known and can be used with theprovided methods.

Also exemplary of the cooling apparatuses are water baths and ice baths,for example, water baths and/or ice baths in which the vessel(s) areplaced, for example, during homogenizing.

Typically, the cooling apparatus can be used to cool the liquid tobetween 25° C. or about 25° C. and 45° C. or about 45° C., for example,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44 or 45° C., typically between 25° C. and 43° C., typically between35° C. and 43° C., for example, 26.5° C. Typically, the cooling is rapidcooling, for example, cooling to between 25° C. or about 25° C. and 45°C. or about 45° C., for example, between 35° C. and 43° C., for example,26.5° C., in between 15 minutes or about 15 minutes and 2 hours or about2 hours, typically, between 30 minutes or about 30 minutes and 60minutes or about 60 minutes, for example, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59 or 60 minutes.

f. Transfer Means

Transfer means are used with the provided methods to transfer liquidfrom one vessel to another vessel, for example, to transfer the contentsof one or more vessels to one or more other vessels, for example, totransfer the water phase to the oil phase vessel (e.g., the oil phasetank) or to transfer the oil phase to the water phase vessel (e.g., thewater phase tank), in order to form the emulsion. Exemplary of theequipment used for the transfer means are transfer pumps and associatedaccessories, for example, ball valves, sanitary fittings (for example,sanitary fittings sold by Granger, Inc., Lake Forrest Ill.) and transferhoses (for example, hoses sold by Sani-Tech West, Oxnard, Calif.), forexample, food grade hoses attached to the transfer pumps. Exemplary ofthe transfer pumps that can be used with the provided methods is theTeel Pump (Model 2P377B), Granger, Inc. Lake Forrest II, a self-primingpump having a power rating of 2 HP, 60 Hz voltage 208-230/460 AC, speedof 3450 rpm. Other pumps, for example, other self-priming pumps fromGrainger, Inc., can be used as part of the transfer means in theprovided methods. Alternatively, transfer means can include means formanually transferring the liquid to another vessel, for example, bypouring, pipetting and/or other well-known methods of manuallytransferring liquids.

g. Evaluation Equipment

Evaluation equipment is used to evaluate one or more properties of thecompositions, for example, the phases of the compositions and/or thefinal concentrates. For example, evaluation equipment can be used tomeasure one or more parameters of the concentrates and/or the phases,for example, the temperature and the pH of the liquids. Exemplary of theevaluation equipment are pH meters and temperature meters. Exemplary ofthe pH/temperature meters is the pH and temperature meter sold by HannaInstruments, (model number HI 8314), which can be used to measure thetemperature and the pH of the mixture(s). Also exemplary of temperaturemeters are temperature probes, for example, digital and/or water-prooftemperature probes, for example, temperature probes sold byCooper-Atkins, Middlefield, Conn., for example, the digital waterprooftemperature probe (Model # DPP400W) from Cooper-Atkins. Other evaluationequipment for evaluating liquids and/or emulsions is well known and canbe used with the provided methods.

3. General Methods for Making the Liquid Nanoemulsion Concentrates

In general, the provided methods for making the concentrates includesteps for generating phases (e.g., oil phase(s) and water phase(s)) andsteps for combining and emulsifying the phases, to form the liquidnanoemulsion concentrates. In some examples, the methods includeadditional steps, such as evaluation, addition of further ingredients,packaging and filtering. The provided methods can be performed using abench-top manufacturing process (typically for small batch sizes).Alternatively, the methods can be performed using a scaled-upmanufacturing process (typically for larger batch sizes). Each of theprovided concentrates can be made using either a scaled-up process or abench-top process. In one example, after the concentrate first is madeusing the bench-top process, the method is scaled up to make largerquantities of the concentrate using the scaled-up process. Whenformulating the concentrates according to the provided methods, theinitial concentrate typically is made by a bench-top method. In oneexample of the formulation methods, a selected formulation then is madeusing a scaled-up process. Any of the concentrates provided herein canbe made with the provided methods, using either manufacturing process.Any method described herein, where the bench-top method is used, can bescaled-up for production of the concentrates using the scaled-upprocess.

Generally, the provided methods for making the liquid nanoemulsionconcentrates include first generation steps, whereby one or more oilphases and one or more water phases are produced. Generation of thewater phase and generation of the oil phase typically are performed inat least two separate vessels, for example, an oil phase vessel and awater phase vessel. Each of the generation steps typically includes amixing step and a heating step, which can be performed simultaneously,sequentially in any order, or partially simultaneously.

To generate the water phase, water phase ingredient(s) (e.g., the polarsolvent (e.g., water, propylene glycol, glycerin or other polar solvent)and, in some examples, additional water phase ingredients) are added toa water phase vessel. The ingredient(s) are mixed, typically using astandard mixer, and heated, for example, using a heating apparatus.Typically, the water phase ingredients are heated to a low heattemperature, for example, to 60° C. or about 60° C. To make the oilphase, the oil phase ingredients (e.g., non-polar compound(s),surfactant(s) and, in some examples, other oil phase ingredient(s)) areadded to an oil phase vessel. The oil phase ingredient(s) are mixed,typically using a standard mixer, and heated, for example, using aheating apparatus. Typically, the ingredients are heated to a low heattemperature, for example, to 60° C. or about 60° C. The mixing/heatingof the water and oil phase can be performed simultaneously orsequentially, in any order. In one example, generation of the oil phaseis performed subsequently to generation of the water phase, for example,to preserve the non-polar active ingredient, for example, to prevent itsoxidation. Typically, both phases are heated to the desired temperature,for example, low heat temperature, and/or until the ingredientsdissolve, prior to combining the oil and the water phases in asubsequent emulsification step.

In general, the methods further include an emulsifying step. For theemulsifying step, the oil and water phases are combined, for example,using one or more transfer means. The oil and water phases areemulsified, typically with mixing, typically homogenizing, for example,using high shear, in order to generate an emulsion (e.g., the liquidnanoemulsion concentrate). The emulsifying step can be performed in thewater phase vessel, the oil phase vessel, or a separate vessel.

Typically, during the emulsifying step, the forming emulsion is cooled,for example, rapidly cooled, for example, using one or more coolingapparatuses. Typically, the cooling step is performed simultaneouslywith the emulsifying step. In one example, the cooling is performeduntil the emulsion reaches a temperature of between 25° C. or about 25°C. and 43° C. or about 43° C.

The provided methods can include additional steps, for example,evaluation steps, steps for adding additional ingredients, purification(e.g., filtration) steps, and/or packaging/holding steps, as detailedbelow.

a. Generating the Water Phase

Typically the water phase ingredients are weighed and/or measured, forexample, using one or more scales (e.g., one or more of the scalesdescribed herein), before addition to the water phase vessel (e.g., anyvessel described herein). In one example, the amount of each ingredientto be added to the water phase vessel is determined according to theprovided methods for formulating the concentrates. Typically, thedesired concentration, by weight (w/w), of the final nanoemulsionconcentrate is used to calculate the amount of each water phaseingredient that is added to the water phase vessel. Alternatively, thedesired volume per weight, volume per volume or weight per volume can beused to calculate the correct amount of an ingredient to be measured andadded to the vessel.

In one example, when water is the polar solvent, impurities in thewater, for example, city water, are removed using one or more purifiers(e.g., one or more purifiers as described herein) above, before addingthe water to the water phase tank. In one example, the water is purifiedby passage through using the following purifiers, sequentially: a carbonfilter, an ion exchange purifier, a reverse osmosis purifier and anend-point filter, for example, a 100 micron end-point filter, beforebeing added to the water phase vessel.

Typically, the water phase ingredient(s) are mixed in the water phasevessel using a standard mixer (e.g., any of the standard mixersdescribed herein) and heated, typically simultaneously or, in part,simultaneously, using a heating apparatus (e.g., any of the heatingapparatuses described herein). Typically, the water phase is heated suchthat the water phase ingredients reach a low heat temperature, forexample, between about between 45° C. or about 45° C. and 85° C. orabout 85° C., for example, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85° C., typically, 60° C.or about 60° C., for example, to prevent oxidation of the non-polaringredients and/or maintain the stability of the ingredients. Typically,mixing and/or heating of water phase ingredients in the water phasevessel is continued, for example, prior to combining the water phase andthe oil phase. In one example, the water phase is mixed and/or heateduntil the water phase ingredients have dissolved. Typically, thetemperature of the water phase is maintained with mixing prior tocombining the oil and water phases.

i. Water Phase Ingredients

The water phase includes one or more polar solvent, such as water,diols, such as propylene glycol and sugar alcohols, such as glycerin,and, in some examples includes other water phase ingredients. Typically,water phase ingredients are hydrophilic and/or amphipathic ingredientsof the liquid nanoemulsion concentrate. For example, oils and otherlipophilic ingredients typically are not added to the water phase.Certain ingredients, for example, ingredients having hydrophobic andhydrophilic moieties, for example, surfactants and co-surfactants, canbe added to either the oil or the water phase, or to the oil and thewater phase. Exemplary water phase ingredients include, but are notlimited to, polar solvents, e.g., water, typically filtered water,propylene glycol, glycerin and other diols; emulsion stabilizers; pHadjusters, for example, phosphoric acid and/or citric acid; flavors;surfactants; co-surfactants, for example, phosphatidylcholine and/orquillaja saponin; and preservatives.

Water phase ingredients can be added to the water phase simultaneouslyand/or sequentially, in a specific order. In one example, one or morewater phase ingredients is added first and heated, prior to addition offurther ingredient(s). In one example, when the water phase ingredientsinclude a polar solvent and an emulsion stabilizer, these ingredientsare added sequentially, in the following order: 1) polar solvent; 2)emulsion stabilizer. In one example, when the water phase ingredientsinclude water and an emulsion stabilizer, these ingredients are addedsequentially, in the following order: 1) water; 2) emulsion stabilizer.In another example, when the water phase ingredients include asurfactant, a polar solvent (e.g., water) and an emulsion stabilizer,these ingredients are added to the water phase vessel sequentially, inthe following order: 1) surfactant; 2) polar solvent (e.g., water); 3)emulsion stabilizer. Alternatively, the water phase ingredients can beadded in any other order. Typically, when the water phase includes asurfactant, particularly when the surfactant is a surfactant that issolid at room temperature, for example, the surfactant is the firstwater phase ingredient added to the water phase vessel. Typically, whenthe water phase ingredients include an emulsion stabilizer, the emulsionstabilizer is the last ingredient added to the water phase vessel.

b. Generating the Oil Phase

Typically the oil phase ingredient(s) are weighed and/or measured, forexample, using one or more scales (e.g., one or more of the scalesdescribed herein), before addition to the oil phase vessel (e.g., any ofthe vessels described herein). In one example, the amount of each oilphase ingredient to be added is determined according to the providedmethods for formulating the concentrates. Typically, the desiredconcentration, by weight (w/w), of the final nanoemulsion concentrate isused to calculate the amount of each oil phase ingredient that should beadded to the oil phase vessel. Alternatively, the volume per weight,volume per volume or weight per volume can be used to calculate thecorrect amount of an ingredient to be measured and added to the vessel.

Typically, the oil phase ingredients are mixed in the oil phase vesselusing a standard mixer (e.g., any of the standard mixers describedherein) and heated, typically simultaneously, using a heating apparatus(e.g., any of the heating apparatuses described herein). Typically, theoil phase is heated such that it reaches a low heat temperature, forexample, between 45° C. or about 45° C. and 85° C. or about 85° C., forexample, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84 or 85° C., typically 60° C. or about 60° C., forexample, to prevent oxidation of the non-polar ingredients and/ormaintain the stability of the ingredients. In one example, one or moreof the oil phase ingredients are mixed and heated according to theprovided methods, prior to addition of the rest of the oil phaseingredients. For example, the non-polar compound can be mixed and heatedwith one or more solvent, for example, an oil, for example, flaxseed oiland/or Vitamin E oil, until the non-polar compound is dissolved in theoil, prior to addition of the other oil ingredients. Typically, the oilphase ingredients are mixed in the oil phase vessel until dissolved.Typically, the temperature of the oil phase is maintained with mixingprior to combining the oil and water phases.

In some examples the oil and/or the water phase can be made in more thanone vessels, for example, by mixing one or more of the oil phaseingredients in one vessel and mixing the one or more other oilingredients in another vessel. In this example, the mixed oil phaseingredients in the separate vessels either can be mixed together priorto emulsifying with the water phase, or alternatively, can be addedseparately, during emulsification, to the water phase.

i. Oil Phase Ingredients

The oil phase includes the non-polar compound, for example, thenon-polar active ingredient and, in some examples, other oil phaseingredients. Typically, oil phase ingredients include one or morelipophilic and/or amphipathic ingredients of the liquid nanoemulsionconcentrate. Oil phase ingredients typically do not include aqueousingredients or hydrophilic ingredients. Certain ingredients, forexample, ingredients having hydrophobic and hydrophilic moieties, forexample, surfactants and co-surfactants, can be added to either the oilor the water phase, or to the oil and the water phase. Exemplary ofingredients used in the oil phase of the provided concentrates arenon-polar compounds, for example, non-polar active ingredients,including any of the non-polar active ingredients provided herein;emulsion stabilizers, pH adjusters, for example, phosphoric acid and/orcitric acid; surfactants; co-surfactants, for example,phosphatidylcholine and/or quillaja saponin; preservatives, and oils,for example, solvents and other oil phase ingredients.

Oil phase ingredients can be added to the oil phase simultaneouslyand/or sequentially, for example, in any order or in a specific order.In one example, one or more oil phase ingredients is added first andheated, prior to addition of further ingredient(s). In one example, whenthe oil phase ingredients include a surfactant, a preservative, asolvent, a co-surfactant, and a non-polar compound, these ingredientsare added sequentially, in the following order: 1) surfactant; 2)preservative; 3) solvent; 4) co-surfactant; 5) non-polar compound; and6) emulsion stabilizer. In another example, when the oil phaseingredients include a surfactant, a preservative and a non-polarcompound, the ingredients are added sequentially, in the followingorder: 1) surfactant; 2) preservative; 3) non-polar compound. In anotherexample, when the oil phase ingredients include a surfactant, apreservative, a non-polar compound and an emulsion stabilizer, theingredients are added sequentially, in the following order: 1)surfactant, 2) preservative; 3) non-polar compound; and 4) emulsionstabilizer. Alternatively, the oil phase ingredients can be added in adifferent order, for example, any order. Two or more oil phaseingredients can be added simultaneously.

Typically, when the oil phase includes a surfactant, particularly whenthe surfactant is a surfactant that is solid at room temperature, thesurfactant is the first oil phase ingredient added to the oil phasevessel. Typically, when the oil phase ingredients include an emulsionstabilizer, the emulsion stabilizer is the last ingredient added to theoil phase vessel. Typically, the non-polar compound either is the lastingredient added to the oil phase vessel, or is added immediately priorto addition of the emulsion stabilizer, which is the last ingredientadded to the oil phase vessel.

c. Combining and Emulsifying the Oil Phase and the Water Phase

Generally, in the provided methods, following the generation of the oilphase and the water phase, the oil and water phases are combined, forexample, using one or more transfer means (e.g., any of the transfermeans described herein). The combined phases are emulsified, forexample, by mixing, for example, homogenizing, to form an emulsion(e.g., the liquid nanoemulsion concentrate). Typically, the phases aremixed during the combining and the emulsifying steps, for example, usinga homogenizer (e.g., any of the homogenizers described herein). In oneexample, the oil and water phases (e.g., the forming emulsion) furtherare cooled, for example, rapidly cooled, during the emulsifying and/orcombining steps.

i. Combining the Oil and Water Phases

In order to emulsify them, the oil and water phases first are combined,typically by transfer, using one or more transfer means (e.g., any ofthe transfer means described herein). In one example, the oil phase istransferred to the water phase vessel. In another example, the waterphase is transferred to the oil phase vessel. In another example, aplurality of oil phases or water phases are transferred to a water phaseor an oil phase vessel. In another example, the water phase(s) and theoil phase(s) are transferred to another vessel, for example, anemulsification vessel.

Any transfer means can be used to combine the phases. For example, anymeans for transferring the contents of one vessel to another vessel asdescribed above, for example, transfer pumps and associated equipment,for example, sanitary fittings, hoses and/or ball valves; and manualtransfer means, for example, pouring and/or pipetting means or otherknown transfer means. In some examples, the phases are kept clean, forexample, sterile during transfer, for example, by using transfer meanswith sanitary fittings and/or combining the phases in a sterileenvironment.

ii. Emulsifying the Oil and Water Phases

Simultaneous to and/or subsequent to the combination of the phases, thephases are mixed (e.g., homogenized), for example, using a homogenizer(e.g., any of the described homogenizer), to form an emulsion.Typically, the emulsifying is performed in the vessel containing thecombined liquids, for example, the oil phase or the water phase vessel.For this emulsifying step, the oil and water phases are mixed, forexample, after the combining step, typically during and after thecombining step, using a mixer that is capable of emulsifying liquids,for example, a homogenizer, for example, a reversible homogenizer.Typically, the liquids are homogenized using the mixer (e.g.,homogenizer) at low speed, for example, low rpm, for example, between850 rpm or about 850 rpm and 1200 rpm or about 1200 rpm, for example,850, 900, 950, 1000, 1050, 1100, 1150 or 1200 rpm. Lower speeds canreduce the incorporation of air into the nanoemulsion. In some examples,the homogenization can be performed at speeds less than 850 rpm, suchas, for example, between 25 or about 25 rpm and 50 rpm or about 50 rpm,for example at or about 30 rpm, to further reduce incorporation of airinto the nanoemulsion.

The liquids typically are mixed, continuously or intermittently, untilthe liquids are emulsified, for example, in a nanoemulsion. In oneexample, the mixing speed is maintained in order to emulsify the oil andwater phases. In one example, the baffle plate of the mixer is adjusted,for example, by moving the baffle plate further down into the mixture orfurther up out of the mixture, to control the type of mixing, forexample, to switch from downward flow to upward flow and vice versa,during mixing of the emulsion. In another example, the homogenizer canbe adjusted to increase or decrease shear or to maintain the shear at aparticular speed. Methods for homogenizing oil and water phases are wellknown and other methods can be used to homogenize the oil and waterphases in the provided methods.

iii. Cooling

Typically, the emulsion is cooled during mixing, for example, by rapidcooling. In one example, the emulsion is cooled to promote stability ofthe emulsion and emulsification of the phases, for example, bypreventing or minimizing oxidization, for example, oxidization of thenon-polar compound. The cooling, for example, rapid cooling, typicallyis performed using one or more cooling apparatuses, for example, any ofthe cooling apparatuses described herein or any known cooling apparatus.In one example, the cooling apparatus is a recirculating cooler. Inanother example, the cooling apparatus is a water bath or an ice bath.In one example, when the apparatus is a recirculating cooler, fluid fromthe vessel being used for the emulsifying step is recirculated throughthe cooler, and then back to the vessel, to rapidly cool and maintainthe temperature of the mixture during mixing. Typically, the formingemulsion is mixed and cooled until the phases are emulsified and thetemperature reaches between 25° C. or about 25° C. and 43° C. or about43° C., for example, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42 or 43° C. Typically, when the cooling is rapidcooling, the temperature is reached in less than 2 hours or about 2hours, typically less than 1 hour or about 1 hour, for example, in atleast between 30 minutes or about 30 minutes and 60 minutes or about 60minutes, for example, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59or 60 minutes.

Once the oil and water phases have been emulsified, thereby forming anemulsion, for example, a liquid nanoemulsion concentrate, the emulsioncan be used, for example, in the provided dilution methods to make aliquid dilution composition, for example, a beverage, containing theconcentrate. Alternatively, one or more additional steps can beperformed before using the concentrate.

d. Additional Steps

Typically, one or more additional steps is carried out, followingemulsifying the phases, prior to use of the concentrate. For example,the emulsion can be evaluated (e.g., by measuring pH and/or temperatureof the concentrate). In another example, one or more additionalingredients can be added to the emulsion. In another example, thenanoemulsion concentrate is transferred to a holding vessel or apackaging vessel, for example, a holding/packaging vessel, for example,a holding/packaging tank. In another example, the nanoemulsion ispurified, for example, filtered, prior to use. In one example, additionof additional ingredients, evaluation and/or purification, can beperformed in the holding/packaging vessel. Other additional steps can beperformed prior to use.

i. Additional Ingredients

In one example, additional ingredients, for example pH adjusters and/orflavors, can be added to the emulsion after it is formed. In oneexample, citric acid and/or phosphoric acid is added to adjust the pH,for example, until the pH reaches a pH between 2.5 and 3.5, typically,between 2.6 or about 2.6 and 3.2 or about 3.2, for example, 2.6, 2.7,2.8, 2.9, 3.0, 3.1, or 3.2. In another example, one or more flavors isadded to the concentrate, for example, to improve the taste and/or smellof the concentrate and/or beverages containing the concentrate. Inanother example, additional polar solvent, e.g., water, can be added tothe emulsion, for example, in the case of evaporation, to bring theconcentrate to the appropriate volume. Other additional ingredients alsocan be added to the emulsion. Typically, the additional ingredients areadded to the vessel containing the emulsion, for example, the waterphase vessel, the oil phase vessel, the emulsion vessel, or anothervessel, for example, a holding/packaging vessel. Typically, the emulsionis mixed (e.g., using any of the described mixers, typically standardmixers), while the additional ingredients are added.

ii. Evaluation of the Concentrate

Typically, the concentrate is evaluated prior to use. Typically, the pHand/or temperature are measured, for example, using a pH and temperaturemeter. In one example, the pH and/or temperature are evaluated afteradditional ingredients have been added. In one example, furtheringredients can be added to adjust the parameters after evaluation.

iii. Filtering the Concentrate

In one example, the concentrate is purified (e.g., with any of thedescribed purifiers), for example, using an end product filter, prior touse of the concentrate, for example, prior to diluting the concentratein an aqueous medium.

4. Bench-Top Process

In one example of the provided methods for making the liquidnanoemulsion concentrates, the steps of the methods are performed usinga bench-top manufacturing process, which is performed on a bench,counter, table or other surface. Typically, the bench-top process isused to make emulsions having relatively smaller volumes than those madewith the scaled-up process, for example, volumes less than 1 L or about1 L or less than 1 gallon or about 1 gallon, for example, less thanabout 500 mL, for example, 1000, 900, 800, 700, 600, 500, 450, 400, 350,300, 250, 200, 150, 100, 50, or less.

For the bench-top process, the equipment typically is sufficientlycompact to be used on a bench top or other similar surface, typicallysufficiently compact to be moved, for example, lifted, by the artisanusing the methods. For example, the vessels, for example, water phasevessels, oil phase vessels, holding vessels, and packaging vesselstypically are bench-top vessels, for example, flasks, beakers, vials,measuring containers, bottles and/or other bench-top containers. In oneexample, the vessels in the bench-top process is a Pyrex® beaker.Typically, the mixers are mixers that can be used in the bench-topvessels, for example, standard mixers, including hand-held mixers, stirrods, stir bars, magnetic mixers and overhead mixers, for example,mechanical and/or electric overhead mixers and/or other mixers that canbe used in the vessels. Exemplary of appropriate bench-top mixers arestandard mixers, for example, standard mixers sold by IKA®, for example,overhead IKA® mixers, for example, model Nos. RW-14 Basic and RE-16S,which are laboratory stirrers and can be used to mix ingredients, forexample, to generate the oil and water phases. Also exemplary ofappropriate bench-top mixers are homogenizers, for example, reversiblehomogenizers, including The Arde Barinco reversible homogenizer, Modelno. CJ-4E, which can be used to emulsify the phases. Typically, theheating apparatuses are those that can be used with the bench-topvessels, for example, hot plates. The cooling apparatuses typically areapparatuses suited for use with the smaller bench-top vessels, forexample, ice baths and/or water baths into which the vessels can beplaced, for example, for rapid cooling. The evaluation means used in thebench-top process, for example, the temperature and/or pH meters,typically are capable of being placed in the bench-top vessels.

Generally, for the bench-top process, the oil phase and water phase aregenerated by mixing and heating in separate bench-top vessels, forexample, flasks, beakers, vials, measuring containers, bottles and/orother bench-top containers. The mixing typically is performed using anappropriate bench-top mixer, for example, a standard mixer, such as ahand-held mixer, stir rod, stir bar, magnetic mixer and/or overheadmixer, for example, the mixer sold by IKA®, for example, overhead IKA®mixers, for example, model Nos. RW-14 Basic and RE-16S, which arelaboratory stirrers. Typically, heating the oil and water phases isperformed using a heating apparatus appropriate to the bench-top method,for example, a heating apparatus that one or more of the vessels can beplaced upon, for example, a hot plate. For combining the oil phase andthe water phase, one or more phases, typically one phase, typically istransferred manually to another vessel, for example, by pouring,pipetting and/or another manual transfer means. For emulsifying the oiland water phases, a reverse homogenizer typically is used. For coolingthe forming emulsion, for example, for rapidly cooling the emulsion, acooling apparatus appropriate to the bench-top method typically is used,for example, a cooling apparatus that the vessel can be placed upon orinside, for example, a water bath or an ice bath.

5. Scaled-Up Manufacturing Processes

The provided methods for making the liquid nanoemulsion concentrates canbe performed using a scaled-up manufacturing process. A scaled-upmanufacturing process typically is used when the liquid nanoemulsionconcentrate being made has a relatively larger volume than a concentratebeing made with the bench-top process, for example, volumes greater than1 L or about 1 L or greater than 1 gallon or about 1 gallon, forexample, greater than about 500 mL, for example, at least 0.5 L, 1 L, 2L, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100,150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 800, 900,1000, or more gallons.

In general, the scaled-up manufacturing processes are performed withequipment that is compatible with these larger volume batches (batchsizes). For example, the vessels used in the scaled up process typicallyare tanks, e.g., water jacketed tanks, which are equipped with waterjackets that can be used as heating apparatuses to heat the oil andwater phase ingredients during generation of the oil and water phases.The water jackets typically are controlled via control panels.Similarly, the transfer means used in the scaled-up process typicallyinclude transfer pumps and associated fittings, for example, ball valvesand hoses. Exemplary of mixers that are used in the scaled-up processare standard mixers (for example, mounted mixers, for example LIGHTNIN®mixers, for example, Model XJC117 (a fixed-mount, gear drive high-flowmixer, and Model ND2. An exemplary scaled-up process is set forth inFIG. 1 and described in this section, below. The provided methods formaking the concentrates can be performed using this exemplary scaled-upprocess, or any variation of the scaled-up process, for example,eliminating one or more steps of the exemplary process, adding one ormore steps according to the provided method, and/or substituting stepsand/or equipment according to the methods provided herein.

FIG. 1 sets forth a an exemplary scaled-up process 100 for making theliquid concentrate. In this example, the polar solvent is water. Thisexemplary scaled-up process includes the following steps:

a. Water Purification

As noted herein, the polar solvent can include water (including purifiedwater) and other polar solvents, e.g., glycerin and propylene glycol. Inthe example illustrated in FIG. 1, the polar solvent is water 101 (e.g.,city water), which is purified before addition to the water phase vesselby passing the water through the following purifiers, sequentially, inthe following order: a carbon filter 105, ion exchange equipment 106,reverse osmosis equipment 107, a 100 micron end-point filter 108, and a50 micron point-of-use filter 109.

b. Generation of the Water Phase and Oil Phase:

As described above, for generation of the water phase, the polar solventand any other water phase ingredients typically are weighed and/ormeasured, and added to the water phase vessel and mixed, using astandard mixer or other mixer, such as a homogenizer or other mixerdescribed herein, and typically heated during mixing, whereby the waterphase is generated with mixing and heating, typically to low heat (e.g.,60° C., 70° C., 71° C.), according to the provided methods. In theexample of the scaled-up manufacturing process set forth in FIG. 1, thewater phase vessel is a water phase tank 103, which is a water-jacketedtank. In the example illustrated in FIG. 1, the water phaseingredient(s) are mixed using a standard mixer 111, for example, aLIGHTNIN® mixer (for example, model no. XJC117, a fixed-mount gear drivehigh-flow mixer), attached to the tank, for example, mounted on the topof the tank. In the example illustrated in FIG. 1, the heating apparatusused to heat the water phase ingredients is the water jacket of thewater-jacketed tank; temperature on the water-jacket is controlled via acontrol panel.

As described above, for generation of the oil phase, the oil phaseingredients typically are weighed and/or measured, and added to the oilphase vessel and mixed, using a standard mixer or other mixer, such as ahomogenizer or other mixer described herein, and typically are heatedduring mixing, whereby the oil phase is generated with mixing andheating, typically to low heat (e.g., 60° C.), according to the providedmethods. In the example of the scaled-up manufacturing process set forthin FIG. 1, the oil phase vessel is a water-jacketed oil phase tank 102.In the example illustrated in FIG. 1, the oil phase ingredients aremixed using a standard mixer 111, for example, a LIGHTNIN® mixer (e.g.,model ND2), attached to the oil phase tank, for example, mounted on thetank. In the example illustrated in FIG. 1, the heating apparatus usedto heat oil phase ingredients is the water jacket of the water-jacketedoil phase tank; temperature on the water-jacket is controlled via acontrol pane.

c. Combining and Emulsifying the Phases

As described herein, once the oil and water phases reach the desiredtemperature (e.g., 60° C., 70° C., 71° C. or other temperature), afteroil phase and/or water phase ingredients have dissolved, and optionallyafter cooling one of the phases, e.g., cooling the water phase to 60° C.according to the provided methods, the oil and water phases arecombined, via transfer, and emulsified, typically via homogenization. Inone example, the transfer is carried out slowly to prevent clumping inthe forming emulsion, such as by stopping the transfer periodicallywhile continuing to mix the emulsion, or by combining the phases slowlywith mixing. In the example of the scaled-up manufacturing processesillustrated in FIG. 1, the combining of phases is effected bytransferring the oil phase to the water phase vessel, via transfer means112, which include a transfer pump (e.g., a Teel pump, model 2P377B,sold by Granger, Inc.), sanitary fittings, transfer hose(s) (e.g., foodgrade hoses sold by Sani-Tech West) and ball valve(s). Alternatively,the water phase can be transferred to the oil phase. In the example setforth in FIG. 1, to begin the combining/emulsifying steps, a homogenizer110 (e.g., an Arde Barinco, Inc. reversible homogenizer), mounted on thewater phase tank, is turned on, for example, at 850-1200 rpm. The ballvalves then are opened and the transfer pump turned on, therebyeffecting transfer of the oil phase liquid to the water phase tank viathe transfer hose(s). As the phases are combined, the mixture ishomogenized by continued mixing with the homogenizer 110.

In some examples of the scaled-up manufacturing process, to preventclumping, the pump is periodically stopped (e.g., by turning off thepump), while continuing to mix with the mixer, during emulsification. Inone aspect of this example, this method to prevent clumping is used whenthe polar solvent is a solvent other than water, such as propyleneglycol or glycerin. During mixing, the homogenizer can be adjusted, forexample, by adjusting the baffle plate on the homogenizer to achieve andmaintain an emulsion, for example, by moving the baffle plate furtherinto the forming emulsion and/or further out of the forming emulsion. Inone example, the shear speed is adjusted to a speed where the oil phasecan be seen coming out the top of the mixer. In one example, thisadjustment is used when the polar solvent is a solvent other than water,such as propylene glycol or glycerin.

d. Cooling

As described herein, the forming emulsion typically is cooled, typicallyrapidly cooled, during the emulsion step. In the scaled-up process, asshown in the example illustrated in FIG. 1, the rapid cooling typicallyis effected by repeatedly passing the forming emulsion throughrecirculating cooler 115 (e.g., Model No. OC-1000 RO, sold by Turmoil,West Swanzey, N.H.), which is attached to the water phase tank.Homogenization continues during the cooling step, for example, atbetween 850 and 1200 rpm. The cooling continues, for example, until thetemperature of the emulsion reaches between at or about 25° C. and at orabout 43° C., such as between at or about 25° C. and at or about 35° C.,between at or about 35° C. and at or about 43° C., or at or about 40° C.Typically, the rapid cooling is carried out for between at or about 30and at or about 60 minutes.

e. Additional Steps

As described herein, additional steps can be performed after theemulsion is formed. In the example of the scaled-up manufacturingprocess set forth in FIG. 1, the additional steps include transferringthe emulsion, via transfer means 112, which include a transfer pump(e.g., a Teel pump, model 2P377B, sold by Granger, Inc.), sanitaryfittings, transfer hose(s) (e.g., food grade hoses sold by Sani-TechWest) and ball valve(s), to a holding/packaging tank 104. Transfer isperformed by turning on the transfer pump and opening the ball valves.Additional ingredients can be added, for example, pH adjusters, forexample, while monitoring pH, sufficient to bring the nanoemulsion to anappropriate pH, for example, between about 2.6 and 3.2. Flavors can alsobe added. The additional ingredients are mixed into the concentrateusing a standard mixer 111. The addition and mixing of additionalingredients, and/or evaluation can be performed in the holding/packagingtank 104; alternatively it can be performed prior to transfer to theholding/packaging tank, for example, in the water phase tank 103.

Variations of this exemplary scaled-up process (FIG. 1) also can beperformed using the provided methods, including any of the variationsdescribed herein, to make the concentrates. For example, by eliminationand/or modification of one or more steps and/or equipment, according tothe general methods provided herein.

D. Methods for Preparing Liquid Dilution Compositions Containing theDiluted Concentrates

Also provided herein are methods for diluting the liquid nanoemulsionconcentrates to make liquid dilution compositions, typically, aqueousliquid dilution compositions, containing the non-polar compounds.Generally, the nanoemulsion concentrate is diluted into an aqueousmedium, for example, a beverage, for example, soda, water milk, juice,fitness drinks, nutritional beverage, nutritional supplement, or otheraqueous food or beverage. The concentrate and the aqueous medium can bemixed, for example, by stirring and/or blending or by any known mixingmeans. The concentrate disperses into the aqueous medium to form anaqueous liquid dilution composition, for example, a clear or partiallyclear aqueous liquid dilution composition. The aqueous liquid dilutioncomposition can be evaluated, for example, to assess the clarity, taste,smell, and/or stability of the liquid.

In one example, the liquid nanoemulsion concentrate is diluted in theaqueous medium, for example, water by heating the aqueous medium, forexample, by heating the aqueous medium, for example, to at least 40° C.or at least about 40° C., for example, 41, 42, 43, 44, 45, 46, 47, 48,49, 50 or more ° C., for example, 48.9° C. In this example, the liquidnanoemulsion concentrate is added, at an appropriate dilution, asdescribed herein, to the heated aqueous medium, and stirred untildispersed or dissolved in the solution. The resulting liquid dilutioncomposition can then be cooled, for example, to room temperature, forexample, 25° C. or about 25° C. Following dilution, the aqueous liquiddilution composition can be packaged, for example, by transferring tocontainers, for example, vials or beverage containers. In one example, aportion of the liquid dilution composition is transferred to vials foranalysis, for example, evaluation of properties, such as clarity,turbidity, taste, smell, ringing, crystal formation and/or otherproperties.

Exemplary of equipment used for diluting the liquid nanoemulsionconcentrates to form the liquid dilution compositions containing thediluted concentrates are beakers, for example, Pyrex® glass beakers, hotplates, for example, the Thermolyne hot plate, model number 846925 ormodel number SP46615, stir rods, temperature meters, for example,temperature probes, for example, Cooper Temperature Probes (model no.DPP400W) and scales, for example, the OHUAS 2.0 Kg scale (Model #CS2000) and/or the Sartorius Analytical Scale (model BA 110S).

1. Dilutions

Typically, the provided concentrates can be diluted into aqueous mediato form aqueous liquid dilution compositions over a wide range ofdilutions. In one example, the concentrate can be diluted so that theaqueous liquid dilution composition contains between 0.05 g or about0.05 g and 10 g or about 10 g, typically between 0.05 g and 5 g, of theliquid concentrate per 8 fluid ounces of the liquid, at least 8 fluidounces of the liquid or less than 8 fluid ounces of the liquid, or persingle serving of the liquid. For example, the concentrate can bediluted so that the aqueous liquid dilution composition contains 0.05 g,0.06 g, 0.07 g, 0.08 g, 0.09 g, 0.1 g, 0.2 g, 0.3 g, 0.4 g, 0.5 g, 0.6g, 0.7 g, 0.8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, org of the concentrate per 8 fluid ounces, about 8 fluid ounces, or atleast 8 fluid ounces or at least about 8 fluid ounces of the aqueousmedium, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,25, 30, 35, 40, 45, 50, 100, 200 or more fluid ounces, of aqueousmedium.

In another example, the concentrate is diluted so that the aqueousliquid dilution composition contains between 1 mL or about 1 mL and 10mL or about 10 mL of the liquid concentrate, for example, 1 mL, 2 mL, 3mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL or 10 mL of the concentrate, per8 fluid ounces, about 8 fluid ounces, at least 8 fluid ounces or atleast about 8 fluid ounces, or less than 8 fluid ounces or less thanabout 8 fluid ounces, or per serving size, of the aqueous medium, forexample 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35,40, 45, 50, 100, 200 or more fluid ounces, of aqueous medium.

In another example, the liquid concentrate is diluted so that theaqueous liquid dilution composition contains at least 10 mg or about 10mg, typically at least 25 mg or about 25 mg, typically at least 35 mg,of the non-polar compound, for example, the non-polar active ingredient,per 8 fluid ounces (0.236588 liters) or about 8 fluid ounces, at least 8fluid ounces or at least about 8 fluid ounces (0.236588 liters) of theaqueous medium, or less than 8 ounces or less than about 8 ounces, orper serving size, of the aqueous medium; for example, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100,110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240,250, 260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450, 475, 500,550, 600, 700, 800, 900, 1000, 1500, 2000 mg, or more, of the non-polarcompound per at least 8 fluid ounces (0.236588 liters) or at least about8 fluid ounces of aqueous medium.

2. Analyzing the Aqueous Liquid Dilution Compositions Containing theLiquid Concentrates

Properties of the aqueous liquid dilution compositions containing theliquid concentrates can be evaluated using a number of differentevaluation means. For example, the clarity; desirability for humanconsumption, for example, pleasant taste, and/or smell, for example,lack of “fishy” taste/smell, lack of “ringing” and lack of crystalformation; stability, for example, lack of oxidation, “ringing,”precipitation and/or visible phase separation, over time; and safety forhuman consumption, can be evaluated. Several of these properties can beevaluated empirically, for example, by observing the liquids immediatelyor over time, or by smelling and/or tasting the liquids. In one example,after evaluation of the aqueous liquid dilution compositions, theconcentrates are re-formulated to adjust one or more parameters. Inanother example, the dilution factor can be adjusted.

a. Clarity/Turbidity

Clarity of the aqueous liquid dilution compositions can be evaluatedusing one or more of several approaches, for example, empiricalobservation, measurement of particle size and/or measurement of aturbidity value. The measurement can be qualitative or quantitative. Inone example, a particular quantitative or qualitative clarity value isspecified. In another example, the clarity of a liquid can be expressedin relation to the clarity of another liquid, for example, an aqueousliquid dilution composition made according to the provided methods, or abeverage, for example, a beverage that does not contain the liquidconcentrate. In this example, the liquid can be as clear as, less clear,or more clear than the other liquid. For example, an aqueous liquiddilution composition containing the liquid concentrate diluted in abeverage can be as clear or about as clear as the same beverage thatdoes not contain the concentrate. Either type of evaluation can be donequalitatively, for example, by empirical evaluation, or quantitatively,for example, by taking a measurement of particle size or turbidity.

i. Empirical Evaluation

In one example, the clarity/turbidity of the aqueous liquid dilutioncomposition is evaluated qualitatively, for example, by observation. Inone example, a liquid is considered clear if it does not have a cloudyappearance and/or if it contains no particles or few particles that areobservable with the naked eye. In another example, the liquid can beconsidered relatively clear or relatively turbid based on comparison toother liquids, for example, water, fruit juice, soda, and/or milk and/orother aqueous liquid dilution composition(s) made according to theprovided methods. For example, the aqueous liquid dilution compositioncan be as clear or about as clear as water or another liquid, forexample, a beverage. For example, the liquid containing the liquidconcentrate diluted in a beverage can be as clear or about as clear asthe beverage that does not contain the liquid concentrate. In a relatedexample, the liquid can be clear or partially clear when there is nosubstantial difference, for example, no observable difference, betweenthe aqueous liquid dilution composition containing the concentrate andthe aqueous medium that does not contain the concentrate. A clear liquidis not necessarily colorless. For example, a yellow liquid that containsno (or few) visible particles or cloudiness can be clear. In anotherexample, the lack of crystal formation or of “ringing” can be indicativeof a clear liquid.

ii. Particle Size

In another example, clarity/turbidity are assessed by quantitativelymeasuring particle size and/or number of particles, in the aqueousliquid dilution composition. In this example, the clarity can beexpressed as a numerical representation of the particle size, or as acomparison to the particle size of another liquid.

Methods for measuring particle size of liquids are well known. Anymethod for measuring particle size can be used, provided that it issensitive to the particle size in the expected and/or appropriate rangesof the provided aqueous liquid dilution compositions. For example,particle size analysis is available commercially, for example, fromDelta Analytical Instruments, Inc., North Huntingdon, Pa. In oneexample, the particle size of the aqueous liquid dilution composition ismeasured, for example, by Delta Analytical Instruments, Inc., using alight-scattering analyzer, for example, a dynamic light scatteringanalyzer, for example, the Horiba® LB-550, which can measure particlesizes within a range of 0.001 micron to 6 micron and uses aFourier-Transform/Iterative Deconvolution technique for reporting dataand can measure sample concentrations from ppm to 40% solids; theHoriba® LA-920, which is a laser light-scattering instrument having anHe—Ne laser and a tungsten lamp that can determine particle sizes from0.02 micron to 2000 micron using Mie Theory; and other analyzersavailable from Delta Analytical Instruments, Inc.

Alternatively, particle size can be measured by viewing the liquid undera microscope under magnification, for example, a 640× magnification.Particle size then can be measured by comparison to a measuringstandard, for example, a ruler, which also is viewed under themagnification. In one example, particles about 25 nm or greater thanabout 25 nm are visible, while particles less than 25 nm are notvisible, for example under a 640× magnification.

iii. Turbidity Measurement

In another example, the clarity/turbidity of the liquid is evaluatedand/or expressed using a turbidity measurement, for example,Nephelometric Turbidity Units (NTU). In this example, turbidity ismeasured optically, to obtain a value indicating the cloudiness orhaziness of the liquid, which correlates with the number and size ofparticles suspended in the liquid. The more clear a liquid is, the lowerits turbidity value. Turbidity can be measured optically, for example,using a nephelometer, an instrument with a light and a detector. Thenephelometer measures turbidity by detecting scattered light resultingfrom exposure of the aqueous liquid dilution composition to an incidentlight. The amount of scattered light correlates with the amount and sizeof particulate matter in liquid, and thus, the clarity. For example, abeam of light will pass through a sample having low turbidity withlittle disturbance, creating very little scattered light, resulting in alow turbidity (NTU) value reading. Other methods for measuring turbiditycan be used, including commercial services for measuring turbidity, forexample, the services available through ACZ Laboratories, Inc.,Steamboat Springs, Colo.

The following examples are included for illustrative purposes only andare not intended to limit the scope of the invention.

E. EXAMPLES Example 1 General Procedure Used to Prepare the LiquidNanoemulsion Concentrates

Tables 2A-2D and 3, below, set forth ingredients that were used to makeexemplary liquid nanoemulsion concentrates, described in further detailin Example 2, according to the provided methods. Each of theseconcentrates contained a non-polar active ingredient, a polar solvent,and a surfactant. Each concentrate further contained a natural,GRAS-certified, preservative (benzyl alcohol), and was produced,according to this general method, in a 1000 gram (g) or 500 g batch(batch sizes indicated in Tables).

Each of Tables 2A-D and 3 sets forth the milligrams (mg) per 2 mLserving of each ingredient in the exemplary concentrate, the percentage,by weight (of the total concentrate), for each ingredient and the amountin grams (g) of each ingredient per 1000 g batch. Also indicated in eachtable, in the “phase” column, is whether each ingredient was added tothe water phase (“water”), the oil phase (“oil”) or was added later, tothe emulsion formed after combining the oil and water phases in theemulsification step (“emulsion”).

Each of the liquid nanoemulsion concentrates set forth in Examples 2 and3 was made using a bench-top process of the provided methods. To makelarger batch sizes, the bench-top process can be scaled up to make anyof these exemplary concentrates in Examples 2 and 3, using a scaled-upmanufacturing process of the provided methods as described herein.

The bench-top process for making the concentrates in Examples 2 and 3was performed using the following general steps (further details areprovided in the individual examples):

To make the concentrates, the indicated amount of each ingredient wasweighed using a Toledo Scale (Model GD13x/USA), Sartorius BasicAnalytical Scale (Model BA110S) or an OHAUS Scale (Model CS2000).Selection of scale(s) depended on the weight of the particularingredient(s).

To generate the water phase, the water phase ingredients (indicated by“water” in each table in the “phase” column), were added, in theindicated amount (g/batch), to a water phase vessel (a Pyrex® beaker),and mixed using a reversible homogenizer (Arde Barinco, Inc.; ModelCJ-4E), at 30 RPM. During mixing, the water phase ingredients wereheated until the ingredients reached the desired temperature of 60° C.,using a hot plate as the heating apparatus (a Thermolyne hot Plate Model# SP46615, Barnstead International, Dubuque, Iowa). The temperature ofthe water phase and speed of mixing was maintained before combining andemulsifying the water and oil phases. A temperature meter (temperatureprobe (Model # DPP400W, Cooper-Atkins)) was used to evaluate (measure)the temperature of the water phase. The water phase ingredients includeda polar solvent (water) and additional water phase ingredients, whereindicated.

The oil phase ingredients (indicated by “oil” in each table in the“phase” column) were added to an oil phase vessel (a Pyrex® beaker), andmixed using a standard mixer (IKA® model No. RE-16 1S, which is anoverhead mixer (laboratory stirrer) compatible with the bench-topprocess). The oil phase ingredients included a non-polar activeingredient and other oil ingredients as indicated in the Examples.

As the oil phase ingredients were mixed, they were heated using a hotplate as a heating apparatus (a Thermolyne hot Plate Model # SP46615,Barnstead International, Dubuque, Iowa), to a desired temperature of 60°C. and generally mixed at this temperature until ingredients haddissolved, and maintained at the temperature before mixing with thewater phase. A temperature meter (temperature probe (Model # DPP400W,Cooper-Atkins)) was used to evaluate (measure) the temperature of theoil phase.

After both phases had reached the appropriate temperatures (60° C.) andthe oil phase components had dissolved, the phases were combined andemulsified. Emulsification was effected with a reversible homogenizer(Arde Barinco, Inc.; Model CJ-4E). The reversible homogenizer, which wasused to mix the water phase ingredients, was maintained at 30 RPM formixing during the emulsification step. While mixing with the homogenizerat this speed, the oil phase was transferred to the water phase vesselby pouring it from the oil phase vessel into the water phase vessel.Mixing with the homogenizer was continued, with adjustment of the baffleplate on the homogenizer to achieve and maintain an emulsion, forexample, by moving the baffle plate further into the forming emulsionand/or out of the forming emulsion. During emulsification, the formingemulsion was rapidly cooled by placing the water phase vessel (beaker)in a water bath, until the temperature of the liquid reached a desiredtemperature, as indicated in the Examples, between 35° C. and 43° C.,(typically, for a time period between about 30 and about 60 minutes).

In some examples, after emulsifying and rapidly cooling, additionalingredients were added, where indicated, in the individualExamples/Tables. In some examples, pH adjusters (e.g., citric acid) wereadded after combining and emulsifying the oil and water phases(indicated by “emulsion” in the phase column) while mixing with thereversible homogenizer (Arde Barinco, Inc.; Model CJ-4E). The pH of theemulsion was measured using a pH and temperature meter (HannaInstruments, model HI 8314). When needed, the pH was adjusted with theappropriate amount of a pH adjuster (amount indicated in tables), forexample, citric acid, until the emulsion reached a pH of between 2.6 and3.4.

As a final step, the concentrates were filtered using a 100 micronend-product filter, before further evaluation, dilution, and/or use.

Example 2 Liquid Nanoemulsion Concentrates with PUFA-ContainingNon-Polar Compounds

This example sets forth the details of exemplary liquid nanoemulsionconcentrates containing non-polar compounds (non-polar activeingredients) containing polyunsaturated fatty acids (PUFAs). ThePUFA-containing non-polar active ingredients in the exemplifiedcompositions were omega-3 fatty acids, omega-6 fatty acids andconjugated fatty acids, including:

A flaxseed oil compound, which was Fresh Flax Oil, obtained fromBarleans Organic Oils, LLC, Ferndale, Wash., which contained not lessthan (NLT) 55% C18:3 alpha-linolenic acid, and was added at an amount of5%, by weight (w/w), of the final concentrate, whereby the concentratecontained 2.5% ALA;

A borage oil compound, obtained from Sanmark LLC, Greensboro, N.C.(Sanmark Limited, Dalian, Liaoning Province, China), which was derivedby pressing and isolating oil from the seeds of Borago officinalis L.This borage oil non-polar active ingredient contained not less than(NLT) 22% C18:3 gamma-linolenic acid (GLA), and was added at an amountof 5%, by weight (w/w), of the final concentrate, whereby theconcentrate contained 1.1% GLA;

A conjugated linoleic acid (CLA) compound, sold under the trade nameTonalin®, by Cognis Corporation, Cincinnati, Ohio, which contained 1.7%,by weight (w/w), C16:0 Palmitic acid: 2.6%, by weight (w/w), C:18Stearic acid, 13.00% C18:1 C9 Oleic acid, 0.20%, by weight (w/w), C18:2C9 C12 Linoleic acid and 81.00%, by weight (w/w), conjugated linoleicacid (CLA), which included 39.70% Conjugated C9, T11 isomer and 39.50%Conjugated T10, C12 isomer. This CLA-containing non-polar activeingredient was added at an amount of 5%, by weight (w/w), of the finalconcentrate; and

Fish oil, containing about 30% DHA/EPA (sold under the name Omega 30 TGFood Grade (Non-GMO) MEG-3™ Fish Oil by Ocean Nutrition Canada Limited,Nova Scotia, Mass.). The fish oil non-polar active ingredient was addedat an amount of 5%, by weight of the final concentrate, whereby theconcentrate contained 1.5% EPA+DHA.

Tables 2A-2D set forth ingredients and other details of liquidnanoemulsion concentrates, each containing one of the PUFA-containingnon-polar compounds described above, the polar solvent water, and asucrose fatty acid ester surfactant. The specific non-polar activeingredient is indicated in each table. The sucrose fatty acid estersurfactant was DK ESTER® F-160, produced by Dai-Ichi Kogyo Seiyaku Co.,Ltd of Japan, and distributed through Montello Inc., Tulsa, Okla. Thewater was city water, which was purified prior to addition to the waterphase vessel, by passage through using the following purifiers,sequentially in the following order: a carbon filter, an ion exchangepurifier, a reverse osmosis purifier and an end-point filter (a 100micron end-point filter).

Each of the concentrates containing these non-polar active ingredientswas made using the general procedure outlined in Example 1 above withthe following details: The water phase was made by adding the purifiedwater, then the sucrose fatty acid ester surfactant, which was dissolvedby mixing and heating to 60° C., which was maintained until combiningwith the oil phase. The oil phase was generated by adding the followingoil phase ingredients to the oil phase vessel, sequentially, in thefollowing order: 1) non-polar active ingredient; 2) preservative; and 3)surfactant, and mixing and heating to a temperature of 60° C., whichalso was maintained until combining.

During emulsification of the oil and water phases as described inExample 1, the emulsion was rapidly cooled as described in the generalmethod, to a temperature of 40° C.

The pH of the emulsion was measured using a pH and temperature meter(Hanna Instruments, model HI 8314) and adjusted with the amount ofcitric acid indicated in the table until the emulsion reached a pH ofbetween 2.6 and 3.4.

TABLE 2A Liquid Nanoemulsion Concentrate with 5% of an DHA-containingNon-Polar Compound (Fish Oil) and 25.2% Sucrose Fatty Acid EstersPercent (by weight) mg/2 mL of Ingredient serving Phase concentrateg/batch Omega 30 TG Food 100 Oil 5 50 Grade (Non-GM) MEG-3 ™ Fish Oil(non-polar active ingredient) Water (polar solvent) 1380.4 Water 69.02690.2 Sucrose Fatty Acid 454 Water 22.7 227 Ester (DK ESTER F-160)(surfactant) Benzyl alcohol 10 Oil 0.5 5 (preservative) Sucrose FattyAcid 50 Oil 2.5 25 Ester (DK ESTER F-160) (surfactant) Citric Acid (pHadjuster) 5.6 Emulsion 0.28 2.8 Totals 2000.000 100.0000 1000

TABLE 2B Liquid Nanoemulsion Concentrate with 5% of an ALA-containingNon-Polar Compound (Flaxseed Oil) and 25.2% Sucrose Fatty Acid EstersPercent mg/2 mL (by weight) of Ingredient serving Phase concentrateg/batch Flaxseed Oil (55% 100 Oil 5 50 Omega-3) (non-polar activeingredient) Water (polar solvent) 1380.4 Water 69.02 690.2 Sucrose FattyAcid 454 Water 22.7 227 Ester (DK ESTER F-160) (surfactant) Benzylalcohol 10 Oil 0.5 5 (preservative) Sucrose Fatty Acid 50 Oil 2.5 25Ester Surfactant (DK ESTER F-160) (surfactant) Citric Acid (pH 5.6Emulsion 0.28 2.8 adjuster) Totals 2000.000 100.0000 1000

TABLE 2C Nanoemulsion Concentrate with 5% of a GLA-containing Non-PolarCompound (Borage Oil) and 25.2% Sucrose Fatty Acid Esters Percent (byweight) mg/2 mL of Ingredient serving Phase concentrate g/batch BorageOil (22% GLA) 100 Oil 5 50 (non-polar active ingredient) Water (polarsolvent) 1380.4 Water 69.02 690.2 Sucrose Fatty Acid 454 Water 22.7 227Ester (DK ESTER F-160) (surfactant) Benzyl alcohol 10 Oil 0.5 5(preservative) Sucrose Fatty Acid 50 Oil 2.5 25 Ester Surfactant (DKESTER F-160) (surfactant) Citric Acid (pH adjuster) 5.6 Emulsion 0.282.8 Totals 2000.000 100.0000 1000

TABLE 2D Liquid Nanoemulsion Concentrate with 5% of a CLA-containingNon-Polar Compound (CLA Oil) and 25.2% Sucrose Fatty Acid Esters Percent(by weight) mg/2 mL of Ingredient serving Phase concentrate g/batchTonalin ® CLA Oil 100 Oil 5 50 (non-polar active ingredient) Water(polar solvent) 1380.4 Water 69.02 690.2 Sucrose Fatty Acid 454 Water22.7 227 Ester (DK ESTER F-160) (surfactant) Benzyl alcohol 10 Oil 0.5 5(preservative) Sucrose Fatty Acid 50 Oil 2.5 25 Ester Surfactant (DKESTER F-160) (surfactant) Citric Acid (pH adjuster) 5.6 Emulsion 0.282.8 Totals 2000.000 100.0000 1000

Example 3 Liquid Nanoemulsion Concentrates with Coenzyme Q-ContainingNon-Polar Compounds

This example sets forth the details of an exemplary liquid nanoemulsionconcentrate made with a non-polar compound (non-polar active ingredient)containing coenzyme-Q, using the general procedure outlined in Example1, above. The non-polar active ingredient in each of these concentrateswas Coenzyme Q10 (CoQ10), sold under the trade name Kaneka Q10™ (USPUbidecarenone) by Kaneka Nutrients, L.P., Pasadena, Tex., containinggreater than 98% ubidecarenone (ubiquinone), and was added at an amountof 5.25%, by weight (w/w), of the final concentrate. In addition to thenon-polar compound, surfactant, polar solvent and preservative (asdescribed in Example 1), the concentrate further contained a non-polarsolvent (Vitamin E oil, sold by ADM Natural Health and Nutrition,Decatur, Ill., under the name Novatol™ 5-67 Vitamin E(D-alpha-Tocopherol; ADM product code 410217)), containing at least67.2% Tocopherol and approximately 32.8% soybean oil); and aco-surfactant (a phosphatidylcholine co-surfactant, sold under the tradename S-100, by Lipoid, LLC, Newark, N.J., derived from soy extract andcontaining greater than 95% phosphatidylcholine).

Table 3 below, sets forth ingredients and other details of a liquidnanoemulsion concentrate containing the CoQ10 non-polar activeingredient, the polar solvent water, and a sucrose fatty acid ester(SFAE) surfactant (the SFAE surfactant sold under the name DK ESTER®F-160 produced by Dai-Ichi Kogyo Seiyaku Co., Ltd of Japan, anddistributed through Montello Inc., Tulsa, Okla.). This concentratefurther contained an emulsion stabilizer (SALADIZER® brand emulsionstabilizer, which was a blend of xanthan gum, guar gum and sodiumalginate) and citric acid (pH adjuster).

The concentrate was made using the general method described in Example 1above with the following details: For the water phase, city water firstwas purified by passage through using the following purifiers,sequentially in the following order: a carbon filter, an ion exchangepurifier, a reverse osmosis purifier and an end-point filter, forexample, a 100 micron end-point filter. The water then was added to thewater phase vessel, followed by the emulsion stabilizer (water phase)and the surfactant. These water phase ingredients were mixed at 30 RMPusing a reversible homogenizer (Arde Barinco, Inc.; Model CJ-4E), whileheating to a temperature of 60° C. This temperature was maintained priorto addition of the oil phase.

For the oil phase, the following oil phase ingredients were added to theoil phase vessel in the following order: 1) non-polar solvent; 2)preservative; 3) co-surfactant, 4) emulsion stabilizer (oil phase) andmixed with the standard mixer and heated to a temperature of 60° C.,until the co-surfactant had dissolved. The CoQ10 non-polar activeingredient then was added and dissolved at 60° C. The amount of theemulsion stabilizer indicated in the oil phase in Table 3 then was addedand dispersed. Temperature was maintained at 60° C. until adding to thewater phase for the emulsion.

During emulsification of the oil and water phase according to Example 1,the mixture was rapidly cooled to a temperature of 40° C. The pH of theemulsion was measured using a pH and temperature meter (HannaInstruments, model HI 8314) and adjusted with the amount of citric acidindicated in the table until the emulsion reached a pH of between 2.6and 3.4.

TABLE 3 Liquid Nanoemulsion Concentrate with 5% Coenzyme Q-containingNon-Polar Compound and 17.75% Sucrose Fatty Acid Esters Percent (byweight) mg/2 mL of Ingredient serving Phase concentrate g/batch VitaminE Oil (5-67) 75.00 Oil 3.750 18.75 (non-polar solvent) SALADIZER ® 1.270Oil 0.06 0.3175 brand emulsion stabilizer (blend of xanthan gum, guargum and sodium alginate) Water (polar solvent) 1430 Water 71.49 357.4SALADIZER ® 6.800 Water 0.34 1.7 brand emulsion stabilizer (blend ofxanthan gum, guar gum and sodium alginate) Phosphatidylcholine 13.4 Oil0.6690 3.35 (Alcolec PC95) (co-surfactant) Sucrose Fatty Acid 355 Water17.7500 88.750 Ester Surfactant (DK ESTER F-160) (surfactant) BenzylAlcohol 10.00 Oil 0.5000 2.5000 (preservative) Kaneka Q10 ™ 105.0 Oil5.2500 26.250 (non-polar active ingredient) Citric Acid 3.800 Emulsion0.1900 0.9500 (pH adjuster) Totals 2000.000 100.000 500

Since modifications will be apparent to those of skill in this art, itis intended that this invention be limited only by the scope of theappended claims.

The invention claimed is:
 1. A liquid nanoemulsion concentrate,comprising: a) a mixture of one or more sucrose fatty acid ester(s) anda PEG-derivative of Vitamin E, wherein: the total amount of sucrosefatty acid ester(s) and the PEG-derivative of Vitamin E is between about16% and about 30%, by weight, of the concentrate; a nanoemulsion is anemulsion in which the particle size of the dispersed droplets is lessthan 1000 nm; and the concentrate contains at least about 1% sucrosefatty acid ester, by weight, of the concentrate, which increasesmiscibility with the polar solvent compared to in the absence of sucrosefatty acid ester; b) a polar solvent in an amount between about 60% andabout 79%, by weight, of the concentrate, and c) a non-polar activeingredient selected from among any one or more of polyunsaturated fattyacids, omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids,Coenzyme Q 10 compounds, and vitamin D3 in an amount between about 5%and about 10%, by weight, of the concentrate.
 2. The liquid nanoemulsionconcentrate of claim 1, wherein the concentrate contains at least about2% sucrose fatty acid ester.
 3. The liquid nanoemulsion concentrate ofclaim 1, wherein the mixture has an HLB value of between about 14 and20, inclusive.
 4. The liquid nanoemulsion concentrate of claim 1,wherein the mixture contains a sucrose fatty acid monoester.
 5. Theliquid nanoemulsion concentrate of claim 1, wherein the mixture containsany one or more of sucrose stearate, sucrose laurate, sucrose palmitate,sucrose oleate, sucrose caprylate, sucrose decanoate, sucrose myristate,sucrose pelargonate, sucrose undecanoate, sucrose tridecanoate, sucrosepentadeconoate, sucrose heptadecanoate and homologs thereof.
 6. Theliquid nanoemulsion concentrate claim 1, wherein the sucrose fatty acidester surfactant contains any one or more of sucrose monostearate,sucrose monolaurate, sucrose monooleate or sucrose monopalmitate.
 7. Theliquid nanoemulsion concentrate of claim 1, wherein the mixture containsat least about 50%, by weight, sucrose monoester.
 8. The liquidnanoemulsion concentrate of claim 1, wherein the mixture contains asucrose fatty acid ester having a carbon chain length of 12, 14, 16 or18 carbons.
 9. The liquid nanoemulsion concentrate of claim 1, whereinthe total amount of sucrose fatty acid esters is between and includingabout 17% and about 26%.
 10. The liquid nanoemulsion concentrate ofclaim 1, wherein the non-polar active ingredient contains at least onepolyunsaturated fatty acid selected from among omega-3 fatty acids,omega-6 fatty acids, vitamin D3 and conjugated fatty acids.
 11. Theliquid nanoemulsion concentrate of claim 10, wherein the non-polaractive ingredient contains a polyunsaturated fatty acid selected fromamong a docosahexaenoic acid (DHA), an eicosapentaenoic acid (EPA), afish oil, a flaxseed oil, a borage oil, an alpha-linolenic acid (ALA), agamma-linolenic acid (GLA), a conjugated linoleic acid (CLA), and a sawpalmetto extract.
 12. The liquid nanoemulsion concentrate of claim 11,wherein the amount of DHA is between and including about 20% and about90%, or between and including about 25% and about 85%; or between andincluding 35% and about 70%, or between and including about 25% andabout 40%, by weight, of the non-polar active ingredient.
 13. The liquidnanoemulsion concentrate of claim 11, wherein the amount of EPA isbetween and including about 5% and about 15%, between and includingabout 5% and about 13%, or between and including about 5% and about 10%by weight of the non-polar active ingredient.
 14. The liquidnanoemulsion concentrate of claim 11, wherein the amount of ALA isbetween and including about 50% and about 80%, or between and includingabout 65% and about 75%, by weight, of the non-polar active ingredient.15. The liquid nanoemulsion concentrate of claim 11, wherein the amountof GLA is at least 22% or about 22%, by weight, of the non-polar activeingredient.
 16. The liquid nanoemulsion concentrate of claim 1, whereinthe polar solvent is a polar protic solvent.
 17. The liquid nanoemulsionconcentrate of claim 16, wherein the polar solvent is selected fromamong water, glycerin, propylene glycol, ethylene glycol, tetraethyleneglycol, triethylene glycol and trimethylene glycol.
 18. The liquidnanoemulsion concentrate of claim 17, wherein the polar solvent is waterand the amount of water is between and including about 65% and about76%, by weight, of the concentrate.
 19. The liquid nanoemulsionconcentrate of claim 1, further comprising a non-polar solvent in anamount sufficient to dissolve the non-polar active ingredient, whereinthe non-polar solvent differs from the non-polar active ingredient. 20.The liquid nanoemulsion concentrate of claim 19, wherein the non-polarsolvent contains a Vitamin E oil, a flaxseed oil, or a combinationthereof.
 21. The liquid nanoemulsion concentrate of claim 19, whereinthe amount of non-polar solvent is between about 1% and about 6%,inclusive, by weight, of the concentrate.
 22. The liquid nanoemulsionconcentrate of claim 1, further comprising an emulsion stabilizer in anamount sufficient to stabilize the concentrate, compared to the absenceof the emulsion stabilizer.
 23. A method for preparing a powder,comprising spray drying or freeze drying the liquid nanoemulsionconcentrate of claim
 1. 24. The liquid nanoemulsion concentrate of claim1, wherein the total amount of sucrose fatty acid esters is between andincluding about 18% and 26%, by weight, of the concentrate.
 25. Theliquid nanoemulsion concentrate of claim 1, wherein the total amount ofsucrose fatty acid esters is between and including about 16% and about18%, by weight, of the concentrate.
 26. The liquid nanoemulsionconcentrate of claim 1, wherein the total amount of sucrose fatty acidesters is at least and including 17% by weight, of the concentrate. 27.The liquid nanoemulsion concentrate of claim 1, wherein the total amountof sucrose fatty acid esters is about 1% to 10%.
 28. The liquidnanoemulsion concentrate of claim 27, wherein the PEG-derivative ofvitamin E is a tocopherol polyethylene glycol diester (TPGD).
 29. Theliquid nanoemulsion concentrate of claim 27, wherein the PEG-derivativeof vitamin E is tocopherol succinate polyethylene glycol (TPGS).
 30. Theliquid nanoemulsion concentrate of claim 1, wherein the amount ofsucrose fatty acid esters is about 1%.
 31. The liquid nanoemulsionconcentrate of claim 1, wherein the PEG-derivative of vitamin E is atocopherol polyethylene glycol diester (TPGD).
 32. The liquidnanoemulsion concentrate of claim 1, wherein the PEG-derivative ofvitamin E is tocopherol succinate polyethylene glycol (TPGS).
 33. Aliquid nanoemulsion concentrate, comprising: a) a surfactant that is amixture comprising a sucrose fatty acid ester (SFAE) and aPEG-derivative of Vitamin E, wherein: the total amount of the surfactantof a) is between about 16% and about 30%, by weight, of the concentrate;and the concentrate contains at least about 1% sucrose fatty acid ester,by weight, of the concentrate, which increases miscibility with thepolar solvent compared to in the absence of sucrose fatty acid ester; b)a polar solvent in an amount between about 60% and about 79%, by weight,of the concentrate, wherein the polar solvent is water; and c) anon-polar active ingredient selected from among any one or more ofpolyunsaturated fatty acids, omega-3 fatty acids, omega-6 fatty acids,conjugated fatty acids, Coenzyme Q10 compounds, and vitamin D3 in anamount between about 5% and about 10%, by weight, of the concentrate.34. The liquid nanoemulsion concentrate of claim 33, wherein thePEG-derivative of vitamin E is a tocopherol polyethylene glycol diester(TPGD).
 35. The liquid nanoemulsion concentrate of claim 33, wherein thePEG-derivative of vitamin E is tocopherol succinate polyethylene glycol(TPGS).
 36. The liquid nanoemulsion concentrate of claim 35, wherein thetotal amount of sucrose fatty acid esters is about 1% to 10%.
 37. Theliquid nanoemulsion concentrate of claim 33, wherein the total amount ofsucrose fatty acid esters is about 1% to 10%.
 38. A method of providingan oil-based additive in a beverage, comprising: adding a liquidnanoemulsion concentrate of claim 1 to an aqueous beverage medium in anamount, whereby the aqueous medium contains an effective amount of thenon-polar active ingredient for supplementation of the diet.
 39. Themethod of claim 38, wherein the beverage is water, soda, milk, juice ora sports nutrition beverage.
 40. The method of claim 27, wherein thenon-polar active ingredient is selected from among coenzyme Q 10, adocosahexaenoic acid (DHA), an eicosapentaenoic acid (EPA), a fish oil,a flaxseed oil, a borage oil, an alpha-linolenic acid (ALA), agamma-linolenic acid (GLA), a conjugated linoleic acid (CLA), and a sawpalmetto extract.
 41. A liquid nanoemulsion concentrate comprising: a) amixture of one or more sucrose fatty acid ester(s) and a PEG-derivativeof Vitamin E, wherein: the total amount of sucrose fatty acid ester(s)and the PEG-derivative of Vitamin E is between about 16% and about 30%,by weight, of the concentrate; and the total amount of sucrose fattyacid esters is at least about 20%, by weight, of the concentrate, whichincreases miscibility with the polar solvent compared to in the absenceof sucrose fatty acid ester; b) a polar solvent in an amount between 60%and about 79%, by weight, of the concentrate, and c) a non-polar activeingredient selected from among any one or more of polyunsaturated fattyacids, omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids,Coenzyme 010 compounds, and vitamin D3 in an amount between about 5% andabout 10%, by weight, of the concentrate.
 42. The liquid nanoemulsionconcentrate of claim 41, wherein the total amount of sucrose fatty acidesters is at least about 23%, by weight, by the concentrate.
 43. Theliquid nanoemulsion concentrate of claim 41, wherein the total amount ofsucrose fatty acid esters is at least about 25%, by weight, by theconcentrate.